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Ocean Acidification: A systems approach to a global problem

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This module is NEWLY being developed and is UNDER CONSTRUCTION.  Please excuse us as we work out needed details. If you would like to see the latest version of this module - please email us for the current dropbox folder (cludwig@systemsbiology.org).

In this curriculum module, students in high school life and marine science courses act as interdisciplinary scientists and delegates to investigate how the changing carbon cycle will affect the oceans along with their integral populations. 

Students closely model what is occurring in laboratories worldwide, and at ISB through Monica Orellana’s research to analyze the effect CO2 has on ocean chemistry, ecosystems and human societies. Students experiment, analyze public data, and prepare for a mock summit to address concerns. Student groups represent key “interest groups”:

  • Developed nations which pollute CO2
  • Marine calcifying organisms which are predicted to suffer dramatically
  • Marine photosynthesizing organisms, specifically diatoms, which may play a role in CO2 sequestering and will likely increase growth in a high CO­­­2 ­environment
  • Developing island nations which largely depend on ecosystem services that will be threatened by ocean acidification

Students design two experiments to observe the effects of CO2 on seawater pH, diatom growth, algal blooms, nutrient availability, and/or shell dissolution.  

Students begin the module by critically assessing different news articles. As a class they combine their findings to develop a network diagram in order to identify the key players (or parts/nodes) in this system.  They use this to plan their next experimental steps.  Students then use inquiry to understand the effects and properties of CO2. They continue by designing a second experiment based on their interest group to further explore how a change in CO2 has impacted their subsystem. Students model collaborative research by designing and completing cohesive sets of experiments that build off others' experiments. In addition to their own data, students use real-time Puget Sound and worldwide ocean and genetic data to predict the response to further disruptions. In the culminating activity, delegates reconvene to present and discuss their findings in reference to the impact on their ocean network. Recommendations are made for scientists, politicians and people across the world. Students reflect on unanswered questions and on what their individual roles in the networks they have studied are, and how they might change their actions in order to positively impact the network.

To download a one page description of this module, please click here.

The diatom Thalassiosira pseudonana. (Scale bar: 1 μm.) (Micrograph courtesy PNAS of Mark Hildebrand, Scripps Institution of Oceanography, San Diego, CA.)


Resources for Teachers - Please email Claudia Ludwig at cludwig@systemsbiology.org or call 206-732-1453 for the most current version of the draft materials.

FUNDING:  This module has been possible through the National Science Foundation (NSF OCE 0928561) and with leveraged dissemination through NIH/NIGMS.  We have also partnered with Sammamish High School in Bellevue, WA and the University of WA to work with high school students in a Project Based Learning Biochemistry class to help build and field test this module. 

Photos:  genome.jgi-psf.org/Thaps3, www.pnas.org/content/105/5/1391/F1.expansion.html

 

Module Adaptations

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The purpose of this page is to showcase the many adaptations teachers have made to our existing systems biology modules.  Some of these adaptations have been made for classrooms, some for summer programs, senior projects, and science fairs.  If you have any module adaptations you would like to share here for others to use, please contact Claudia Ludwig (cludwig@systemsbiology.org) for more information.

Updates for Summer Courses

Exploring Biological Systems and Genetic Research (aka Dynamic DNA)

  • Please see the above link for more information on this 90 hour course provided through a collaboration between Northwest Association for Biomedical Research, DigiPen Institute for Technology, and the Baliga Lab.  The WA Network for Innovative Careers provided 0.5 occupational education credits to eligible10th-12th grade students who attended this 3 week course at DigiPen Institute for Technology.

 

Updates When Working on Science Fairs and Independent Projects

 

Updates by Classroom Teachers for their Classroom  

Eric Kessler, Bioscience Strand Leader for the Blue Valley School District, has adapted Lessons 1-2 from Ecological Networks and the Environmental Impact on Gene Networks module for his bioscience students

  • Lesson 1 and 2 from Ecological Networks Adapted for Biology Students (link coming soon)
  • Adaptation of Environmental Impact of Gene Networks Part I (link coming soon)
  • Adaptation of Environmental Impact of Gene Networks Part II - to accompany the article in American Scientist - May-June 2007 - Extreme Microbes: Salt-loving microorganisms are helping biologists understand the unifying features of life and molecular secrets of survival under extreme conditions by Shil DeSarma.
  • Adaptation of Environmental Impact of Gene Networks Part III (link coming soon)

Amanda Cope at the Monroe, WALeaders in Learning Program combined Ecological Networks and Environmental Impact on Gene Networks into a three week, intensive systems and inquiry and short course.

Since a formal lab setting were not available, students completed all experiments on tables with stir plates and other needed equipment. The lessons were taught in the order listed on our module pages and with Ecological Networks being taught first.  Students creatively designed their own experiments in the Environmental Impact on Gene Networks in teams of 3-5 students - for many this was their first lab experiment.  Amanda and her students made the most of their environment and available resources to make this a very valuable experience for all.

Lesson 4 - Analysis of Lab Results to Verify Network Interactions

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Lesson Title: Lesson 4- Analysis of Laboratory Results to Verify Network Interactions 


 

Objectives: 
What students learn…
  • Scientists use computer programs and simulations to analyze networks because networks are often very complex.
  • One change in the environment could affect an entire cellular network due to the interdependent nature of networks.
  • A centrifuge separates a culture into a supernatant and pellet, which allows the phenotype of cells to be more easily viewed.
  • Qualitative measurement is a way of describing data with words, whereas quantitative measurement is a way of describing data with numbers.
  • Mutant strains, with known genotypes, are a useful tool when studying organisms.
 What students do…
  • Students use wild type and mutant strains of Halobacterium salinarum to investigate quantitatively and qualitatively.
  • Students use a centrifuge to spin down cells as a way to determine the phenotype of cells.
  • Students compare, contrast and critique divergent results from their investigations and discuss/identify possible errors/sources of variation.  
  • Students use a simulation to verify and/or correct their network understanding.
  • Students plan the steps needed for scientists to determine and verify a question using systems methods.
  • Students analyze the role the environment has on gene expression. In this analysis, recognize the components, structure, and organization of systems and the interconnections within and among them.

Instructional Activities: (Approximately three 50-min periods)

Before beginning lesson:         

Teachers  spin mutant broth samples into pellets at 6000 rpm for 15 minutes, pour off supernatant.
Pellet should be easily seen when given to students. Tip: cells will begin to dislodge and go into solution if not spun for 15 minutes (adjust time if spinning at higher rpm). There should be enough wild type and mutant samples to make a set for each team (8 teams per kit.)
This set of activities may play out differently depending on your facilities and equipment. We put the Mutants PowerPoint, Data Collection from the Student Experiment, and Simulation all in one part. They can be done in nearly any order and can happen somewhat simultaneously (esp. gathering O.D. data). Our description below is the way we think would be optimum. Please adjust to meet your needs.

1. Mutants PowerPoint(Lesson 4.ppt)       

a. Entry task: Have students entering room guess how the purple broth (screen slide 2) came to be. “How could we make this in our lab?” Discuss – accept all reasonable guesses (light, oxygen, enzymes). They don’t need to get to the perfect answers at this point (but at least close). [note for teacher: when bat is over expressed it causes halo to makes lots of bR, even without light.]
 
b. Slides 3 and 4. Explain to students what mutants are (forcibly expressed or removed genes) [note to teacher: this is a fairly easy/common procedure in labs now] and ask what Halobacterium mutants (bat +, bat -, bop-) would look like. They can use their networks (and the ppt. slide) to predict the color of these mutants. (have students record phenotypic predictions for each mutant in their notebook.)
 

c. Show the class the mutant pellets and this short (1/2 page) handout for

Have students record the color of the mutants and compare them to their predictions. Discuss any significant differences between predictions and the actual colors of the mutants. 

2. Collect Data from Student Experiments (This started in lesson 2)

Have students record the O.D. of their samples and spin down a pellet and record the color.

Color cubes.doc

 (Depending on number of spectrophotometers and groups this may take a while. To keep the samples comparable the entire classes’ experiments should all be removed from their heat source at nearly the same time. They can be left on the bench for up to 24 hours without much noticeable difference. If it will be any longer before they can take their O.D. the bacteria need to be refrigerated to keep them from continuing to replicate.) 

3. Simulation:  PM Network simulation ws &    PM simulation ws answers

Have students manipulate the simulation to verify their thinking about this network and how their data and the class data and the mutant data fit. (This is a good activity while waiting for O.D. data collection.)  *Teacher note: simulation requires JAVA, try a different browser if it does not load properly.

4. Compile Class Data

Help students gather the class data onto an excel spreadsheet. (There are multiple sheets for each template.)

Summary Data Template.xls

This is a good place to discuss how we took quantitative data and qualitative data (using the color cubes) and what the difference is and how to compare qualitative data to other qualitative data.

5. Analyze Class Data

Have students decide what the best way is to present the data they have gathered as a class. 

Light Dark OD Template.xls

Talk students through the data analysis process. It will be much easier if you decide to have the class work on only the light/dark data as a group. If your class worked on many different experiments, it might be hard to analyze it all together.
          a. analysis is not deciding what the data means, it is deciding what format will work best to show others the data.
          b. the class takes the raw data and turns it into summary data. This data is “prettier” and easy for others to look at and understand.
          c. sometimes it includes averages or trends or graphs

6. Student Review Sheet Lab Review and AnalysisLab Review and Analysis-Answers

          a. the first section scaffolds the students through understanding their experimental data.
          b. the second section adds the information on mutants.
          c. the third section is a set of questions with a graph on light/dark and oxygen/no oxygen data.

7. Teacher prompts for discussions of information gathered from the network, the student experiments, and the mutants:

a. Compare the mutants (known) with student results (unknowns). Did the unknowns turn out as expected?
b. Phenotype = gene expression. Everyone in class has Halobacterium with the same genotype (clones), how can they have different phenotypes? What caused this? What causes this in nature?

c. Review the pictures from the introductory powerpoint: 

Environmental Influence on Gene Networks Introduction

Use the pictures that ask “Why?” (in the beginning only). Do the students have a better suggestion now to answer the “Whys”?
 
d. Now that we have a working network and some examples of mutants that validate our network, what is the next step? What do scientists do in this situation? (They present to their peers before publication) Then what do they do? (Scientists will look at the next question that comes up from their findings [ex. what network does Halobacterium use for energy when bacteriorhodopsin (bR) is not being produced or when there is a lot of Oxygen.])

8. Presentation

Student groups prepare a quick presentation of their findings and how they relate to the network they created (PowerPoint, brochure, document camera, oral presentation) include their data, class data, and the mutants. Each teacher can decide what format and how much time to spend on this part. Included is a sample of directions for a Powerpoint presentation. Halo Response to Environment PPT Format.doc & Halo Presentation Scoring Guide.doc


Assessment:  How will I know they know...

Post assessment (Post Assessment.doc & Post Assessment w-Answers.doc
Presentation of data

Please help us to make improvements to this module.  We would appreciate a few minutes of your time to take a short survey.   TEACHER  MODULE EVALUATION SURVEY and/or STUDENT MODULE EVALUATION SURVEY


Resources

Lesson 4.ppt      

Color cubes.doc

PM Network simulation ws

PM simulation ws answers

Summary Data Template.xls

Light Dark OD Template.xls

Lab Review and Analysis Worsheet

Lab Review and Analysis Answers

Environmental Influence on Gene Networks Introduction.ppt

E.I.G.N. Intro PPT Script

Halo Response to Environment PPT Format.doc

Halo Presentation Scoring Guide.doc

Post Assessment.doc

Post Assessment w-Answers.doc

Please help us to make improvements to this module.  We would appreciate a few minutes of your time to take a short survey.   TEACHER  MODULE EVALUATION SURVEY and/or STUDENT MODULE EVALUATION SURVEY


Accommodations: Students may have trouble deciphering the class data.  Have students seminar in smaller groups before having a whole-class discussion. 


Extension Activities:  Students engage in a virtual lab experience involving transgenic flies.  Focus is on experimental design and what scientist learn from mutation.  Go to http://www.hhmi.org/biointeractive/vlabs/transgenic_fly/index.html

 Short animations illustrating how gene expression results in phenotype changes.  http://www.hhmi.org/biointeractive/evolution/animations.html       

Students use bioinformatics (CLUSTALW and BOXSHADE) to explore why humans do not produce vitamin C (as most mammals do).  This three-part lesson series is called "Psuedogene Suite" and can be found here (the bioinformatics portion is in part 3): http://www.indiana.edu/~ensiweb/bioinfo.html

Lesson 3 - Data Analysis to Propose Network Function

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Unit: Genetics and Heredity

Part: Environmental Influence on Gene Networks


Lesson Title: Lesson 3- Data Analysis to Propose Network Function

Objectives:

What students learn…

  • Cellular networks allow cells to respond dynamically to stimuli.
  • A response to a stimulus occurs by changing the relative amount or function of individual nodes in a cellular network.

  • A stimulus affecting one node can cause a change in many other nodes because they are all interconnected in the network.

  • Some nodes have bigger system-wide affect than others.

  • Understanding relative importance of nodes allows greater understanding of how to perturb and use the system.

  • Multiple data sets from various experimental techniques are needed to show network relationships.

What students do…

  • Students analyze three different data sets to determine the bacteriorhodopsin network and to determine the importance of compiling multiple sets of data.

 


 
Purposes of each element in the lesson
 
Introductory PowerPoint: Background and review to provide context. Students don’t need to memorize details and do not need to be tested later, but they DO need to understand enough of the context to make sense of the data. A mode of providing the info other than PowerPoint or material for students to reference may be better.  (Printable script/description)
 
Jigsaw Data Analysis: Students will jigsaw the three data sets (Homology, Gene Expression/Microarray and Metabolism) and when completed will have created a model for how Bacteriorhodopsin is created and regulated in Halo using light.  (See 'Extension' for an activity involving microarray interpretation.)  See 'Resources' for additional teacher information.
 
Class Discussion of Jigsaw: Teacher will lead discussion on the answer to overarching questions:
          How do Halobacterium cells control the amount of BR expressed in response to light? What is the gene and protein network that regulates the expression of BR?
Teacher should also focus on how this network is a well-understood example of how an environmental change can set off a cascade of events which can change the phenotype (in this case color) of an organism.
 
Instructional Activities: (Two 50 min periods)
Note: This lesson should be done while the Lesson 2 samples are incubating. Students will get their data for Lesson 2 at the beginning of Lesson 4 (unless your equipment limitations require a different schedule – see Advanced Prep for Module document for timing).
 
Introduction:
1.    Cellular networks and systems biology have been pre-assessed, if your students need some support in this area – it can be inserted here.
·         If you want a full network lesson – Lesson 1 & Lesson 2 from the Ecological Networks Module can be used.
·         If you want a quick review (in worksheet form) – use Sample Network Review Worksheet.doc for possible worksheet.
·         If you want just a quick demo for review - You may just be able to use the Cell Phone simulation.
2.    Students may need to review enzyme-substrate information prior to this lesson. This is the point where you can review network interactions/analysis with your students if your students struggled with this on the module pre-assessment. If you have not yet covered enzymes, this could also be inserted here.
3.   The PowerPoint (Connecting bR lab data to other data) leads a discussion on reasons for this lesson (looking at additional data) and how that data can help explain what students will need to consider when analyzing their data.  (Printable script)

Slide 1 – Asks the question...How will you know what your lab results mean? Students will receive population and phenotype data, so they can support or refute their narrow hypothesis. What is important for this lesson is that they know this is not where an experiment ends.

°   Prompt: Once you have supported/refuted your hypothesis – what is next?
If the students say – “Do another experiment” – ask: how do you know your first experiment makes sense (what if your data is totally invalid)? How do you know what ‘next experiment’ to do?  If students say “Repeat it”, ask   what happens if the data is the same….or different.  Again, how do you know your first experiment makes sense (what if your data is totally invalid)?

°   Get students to the idea that once an experiment is complete, scientists will then see how their data fits into the most current understanding of the organism. Some groups may even collaborate by comparing data and results.

Slide 2 – Asks, what did you measure?  Answers should be:  Color and Population Size
Asks, What are some things the data requires you to understand? (What is needed in order to make sense of the data?)   This can go in many directions, but the teacher should remind students of what they know about color (caused by a pigment used in energy production) and cell division (requires energy).

Slide 3 – Asks, can difference in color be due to genes? Ask students for examples.  *Be sure you discuss not just gene variation which can lead to differences in color (like siblings having different hair color), but also the status of those genes (their regulation) can change the phenotype (remind them of the tan line or hydrangea example from Lesson 1 Intro.

°   Click – a diagram pops up to show how, in the presence of a certain enzyme, a colorless compound is converted into a red pigment molecule – giving a pink flower. Remind students of how the proteins in an organism relate to its physical appearance and how a lot of this action is an indirect relationship. This is especially true of enzymes that allow biological reactions to occur.  If students don’t seem to understand, remind them that this DNA is in every other cell of the plant, so... Why are only parts of the petal turning pink and no other portions of the plant?   Answer – in the red parts of the petal the gene making this enzyme is activated (turned on and actively transcribing), in other areas it is off. The “environment” of those cells flip on the enzyme making genes.
*POINT:  homology data (scientist compare proteins of known pathways to see if the ones they are studying are similar) may help determine what is involved in gene expression resulting in a color change.

Next… What might help explain population growth?

Slide 4-6 - population (growth due to cell division) is connected to energy.  Students should learn about current research on pathways of energy production (metabolism data) to help explain their data.
°   During the jigsaw activity, students will look at an energy formation pathway (halo use bR to produce ATP using light energy in Halo which will allow them to get a better idea of whether their results “make sense”.

Slide 7 - shows the GSL and asks “what makes halo purple?”  This is an opportunity or remind students that they are experimenting with both color and population growth so the jigsaw exercise will offer 3 different types of data that could help them answer confidently.

Slides 8 and 9 - shows a diagram of halo’s membrane with purple protein embedded.  Click and a box shows around 1 of the purple proteins.  Click again and it enlarges.

Slide 10 - asks what the purple protein is actually made of.  

Slide 11 - shows it is called bacteriorhodopsin (bR).

Slide 12 - shows retinal and bop inside the cell.  Click and they move together to form bR.  

Slide 13 - asks what purpose of bR.  Allow students to answer.

Slides 14 and 15 - answer (converts light…).  Click and light is added.  Click again and box appears around 1 of the purple proteins. Click again and it enlarges.  * Note that light does not become bR, light increases the amount of BR.

Slides 16 and 17 - click and 2 ATP are made.

Slide 18 - diagram compares ATP production when light is present and absent.

Slide 19 - point out halo in light (left side) has many more purple proteins in membrane than halo in the dark (right side).

Slide 20 - by now students may ask (hopefully) about why there would be more bR in the membrane when light is present.  If they haven’t arrived at this, the slide 20 asks it for them.

Slides 21 to 23 - illustrate the answer (increases gene expression..initiates transcription).  Students will see this during the jigsaw exercise in more detail.    

Slide 24 - the final slide states the guiding question for the lesson: “How do halobacterium cells change the amount of bR in response to light?”  Explain to the students that they will be modeling what is done in science today (especially when studying a whole system).  In setting up their lab they explored environmental influences on halo.  In order to fit their research into what is understood about halo – they must look at data from other experiments/scientists.  Most scientists tend to specialize (and other scientists will be working on related experiments) so they won’t be producing all the data on an organism/system.  During the jigsaw each of the 3 data groups will be acting as a separate lab group. When they get together to share their ideas, that will illustrate how scientists work together across many fields to complete the “picture” of a system (e.g. this data would have come from geneticists and biochemists).

Jigsaw Groups
7.     Explain that in order to understand how halobacteria regulates the production of bR, various data must be obtained, analyzed and combined to get a complete picture of how the network functions. This is the way modern science functions. Tell students that each person will be responsible for analyzing one of three sets of data and they will then share their findings with a small group. Emphasize to students that there may be names of molecules or genes that they are not familiar with. They will not have to memorize the names of these. The teacher may want to give a brief explanation of naming genes (the names could be anything…).
8.     Divide your class into teams of three (the number of groups depends on the number of students in your class). Assign each student in the team as responsible for a different set of data: homology, microarray (gene expression), or metabolism.
9.     You can either have members of the team work individually on their data sets and questions or you could have members split up and meet with other students who are responsible for the same set of data (groups of 2-3 may be appropriate).  It may be helpful for students to work with others analyzing the same data to discuss the data and the questions.
10.   Each group receives three documents, background information about their technology, the data set obtained from their technology, and the student worksheet for each technology. Students should work in pencil as they might need to make changes to their work.
 
       *See 'Resources' at the end for condensed printables.
a.      Microarray data:
2.      Microarray Data
b.      Homology data:
2.      Homology Data
4.      (key for homology)
c.      Metabolic data:
2.      Metabolic data
11.   Students should work on completing the worksheet for ~15-20 minutes. Once students are finished, they should return to their original group. They may need to help others in their group if they are still working.
12.   Have students return to their original team and share their information. Give each group the Group Handout to complete together.  (Group handout answer key.)
·         Common errors:
a.    Check edge used for BOP + Retinal -> BR. Students tend to write two arrows that both lead to BR. This is not really a correct representation of the relationship, since removal of either product will halt BR production. Be sure students understand the difference between the edge used in the PPT (two arrows into 1) and two separate arrows both leading to BR.
b.    Some students may have put their enzymes in nodes. Feel free to refer them back to their Homology worksheet which does not use nodes. Those enzymes describe the edge, they are not a step the reactant “moves through” in the process of becoming the product. They also are not things we are linking together to show a relationship.
 
Class Discussion
13.    After students have completed the network and answered the questions, use the BR network group debrief PowerPoint to scaffold the summative discussion. This shows the network as it should be drawn by the students, showing the information that is added piece by piece. Play the slideshow or some animation will not work.    Script for ppt with details:  (printable script).


Slides 1-6 reviews the influence of light in making bR and reiterates the 2 guiding questions.

Slides 7-12  actual metabolic network is shown.

Slides 13-15 adds in homology data to the metabolic network. 


Slides 16-18 shows metabolic network (again) and focuses on the genes and proteins that would be involved in a microarray.

*At this point, students have seen the result of the 3 data sets and can compare to their own understanding.  The next slides ask students to make predictions on how various changes in the network will affect the outcome.

Slides 19 - 24  asks ‘which genes are affected by bat’. 

*You may wish to point out that enzymes are positively affected (these are the enzymes that become larger in the PowerPoint; when bat is over expressed, these are also over expressed). The effect of when bat is knocked out is then shown. If any enzymes became larger (they actually don’t) they would be ones that are negatively influenced by bat (when bat decreases, the expression of these enzymes increases).  This is a good place to cover a Washington state standard on feedback and systems. This concept can be difficult for students, particularly the idea that a correlated decrease in two nodes signifies a positive relationship. For example, the decrease in CrtB1 when bat is knocked out means the relationship between the two is positive.

Slides 25 - 26 offer a quick comparison of knocked out and overexpressed bat (again).

Slides 27 - 30  students are asked to predict how bat affects the amount of bR produced.  Click and the metabolism network is added. Click to see this network when bat is knocked out.  Click again to show overexpression of bat.  Click to see them at the same time.  Allow students to study this slide.

Slide 31 reminds students of the 2 guiding questions (see slide 6).

Slide 32 shows the network set up.  Ask students to predict what will happen as light is added.  Ask them to closely watch the bR and ATP.  Click and an arrow goes from light to bat.  Click again and arrows go from bat to CrtB1, brp, and bop.  Click again to increase bat; click to show that CrtB1, brp, and bop increase and then click and wait as the network increases from GG-PP to ATP.

*Ask students the mechanism of how bat is influenced by light. What is the exact role of bat?   Answer: Bat increases transcription of genes associated with the formation of bacteriorhodopsin (handout calls it a Transcription Regulator). When light is present, it is able to bind to the UAS region more efficiently (due to a shape change), increasing transcription of these genes. Due to the cascade of reactions, increasing bop and the enzymes leads to an increase in BR production. There are two slides that will illustrate this point with an animation.
(Homology group students should be best able to answer this question.)
*Make sure you get whole class to understand bats role in transcription of the other genes.

Slide 33 shows the network.  Ask students to predict what will happen if light is absent.  Click- light fades. Click-arrows show bat influences CrtB1, brp, and bop.  Click and bat gets tiny. Click and then so does CrtB1, brp, and bop.  Click to watch the nodes in the network change.

Note: See the bR Network Simulation to understand that oxygen does not have the same relationship with bat as light. Oxygen negatively impacts the binding of bat which results in a decrease in BR if oxygen is high.

Slide 34   Point out to students that they have determined the network for producing bacteriorhodopsin. Note the + signs are included to show bat has a positive relationship with CrtB1, brp, and bop in the presence of light.

This slide also asks about other possibilities.  Students can hypothesize when they think a fermentation pathway would be “on”.  From their research students may remember that this pathway for energy production is used when there is no light (and no oxygen). At least students may bring up the possibility of a network that operates in the dark.  *halobacteria can produce ATP using fermentation.

Slides 35-36   shows the bR network in the presence of light.  Click to add the fermentation pathway.

This bottom pathway (arginine fermentation) is currently hypothesized to be another way that halobacteria can make ATP. Alternative pathways allow halobacteria to survive in a variety of different environments (extremophile).

Note: this network does not include ATP gained through the aerobic cellular respiration pathway. To add oxygen’s influence on bat and include this pathway would make this diagram much more complex. For simplicity it has been left off (additionally the full interaction of all the pathways and their regulation is still be studied).

*Just know (as simulation shows) - in high oxygen conditions, the light pathway is off since most ATP is being produced by the more efficient aerobic respiration pathway.

Click to remove the metabolites.  Click again to show the positive or negative relationship bat has with both networks in the presence of light.

Slides 37-43   the network interacts.

The slides will first show the relationship bat has with certain proteins in the BR pathway AND how this same gene has a negative relationship with proteins in the fermentation pathway. The animation goes on to show when bat is over-expressed (the bacteriorhodopsin pathway is active) and when bat is under-expressed (the arginine fermentation pathway is active). The animation will continue to switch through both bat expressions.

Note: You may need to remind students the difference between a positive and a negative relationship. If bat increases, anything with a positive relationship with bat will also increase, while something with a negative relationship would decrease. The reverse is true if bat decreases (+ would also decrease, - would increase).

Slide 44 asks the class why the cell goes through all the trouble of this complicated network? Why is the bR network regulated? Why doesn’t the cell express all of the proteins at the same time?  

This corresponds with the Big Ideas questions you may need to prompt students for them to see how regulated bop would be different than the regulation using bat. This should show students that bat cuts the pathway off at the start, while bop regulation would allow all metabolites to be produced (stops pathway at end). It is wasteful for the cell to produce all proteins all the time. The cell produces energy in the most efficient way. Cells can regulate (turn on or off) genes depending on what proteins are needed by a particular cell. The pathway that halobacteria uses (photo vs. arginine fermentation) depends on the environmental conditions and the most efficient way to produce ATP.

Slide 45   asks students to think about how we could test the model.  

Emphasize that the final constructed network is a hypothesis or model of what we think is happening in the network based on the data.

The students should come up with the light/dark experiment that they just did. They should recognize the limitation of this experiment (can’t tell for sure involvement of bat). Students that had microarray may bring up the idea of a mutant organism (one that has over or under-expression of the bat gene). If students don’t get there, that is fine. Basically, the main point of this question is for students to realize that even after looking at all this scientific data, they still would need to do experiments to show how the whole network interacts together in response to environmental changes (i.e. how systems biology is studied).

Bring the class back to the experiment and talk about how this energy network could be used to analyze experiment results. For classes that have not yet collected their data, a good formative assessment could be to ask each lab group to predict the phenotype of their experiment (at this point the answers should just be purple (lots of BR) or less purple/pink – based on the color of the Halo when the students start). They may also be able to predict whether their sample should have had minimal, average or a lot of growth. If asked, feel free to remind students that cells need energy to divide. So cells that have lots of energy (using oxygen to perform cellular respiration) will have lots of growth, followed next by those with light but no oxygen and lastly those samples that should only be performing fermentation (no light AND no oxygen).

 
 

Assessment: How will I know they know…
 
Students’ hand drawn network of the bR network and answers to analysis questions through oral discussion.
Students’ hypotheses for how the network regulates the production of bR.
 

Resources:
 

Printable ppt script-connect lab ppt

Excellent compilation of animations, movies, resources

Accommodations:
Students who struggle with scientific vocabulary may experience difficulty with the data comparison jigsaw.  Assign data set and group students so that each student's strength is utilized.  'Microarray data' is the most vocabulary challenging, 'Homology data' requires organizational skills and vocabulary, and 'Metabolism data' while dealing with biochemistry is the least vocabulary challenging and the simplest to decipher.
 
Students may benefit from powerpoint slides being printed as handouts with lines for notes.
 

Extension Activities:
A 'wet-lab' simulation to show students how microarrays work,  
kit from Carolina Biological complete with a Teacher Guide and Animation.  Thanks go to A. Malcolm Campbell and colleagues from Davidson College.
 
A 'dry-lab' simulation to show how microarrays work.  Teacher guide,  student information, student data. (Thanks go to Seattle BioMed's education program director, Theresa Britschgi and to EMBL.)
 
Scientists expand their understanding of gene expression by 'knocking out' a gene to determine how it affects a network.  By removing a gene (sap102) thought to be involved in learning and memory, scientists determined that it is indeed, active.  Students are given some background information by way of video and animation and then can see the experiment used and data collected to support this conclusion. (Adapted materials from Dolan DNA Learning Center at Cold Spring Harbor Laboratory.)
 
Scientists compare DNA or amino acid sequences of organisms to determine how closely related they are.  This site is a good, quick review of homology and bioinformatics involving the same model organisms seen in the previous extension.  (Adapted materials from Dolan DNA Learning Center at Cold Spring Harbor Laboratory)
 
 
 
 
 

Lesson 2 - Response of Halo in Different Environmental Conditions

Posted in
Unit: Genetics and Heredity
Part: Environmental Influence on Gene Networks

Lesson Title: Lesson 2- Growth and Phenotypic Response of Halobacterium in Different Environmental Conditions

Objectives:
What students learn…
  • Observations lead to hypotheses and experimental design.
  • Halobacterium (Halo) respond to the amount of light in their environment.
  • Halo change color from pink to purple in the presence of light and from pink to tan/white in the absence of light.
  • Halo grow better in the presence of light.
  • Using a centrifuge separates a culture into a supernatant and pellet, which allows the phenotype of cells to be more easily viewed.
  • Qualitative measurement is a way of describing data with words, whereas quantitative measurement is a way of describing data with numbers.
  • The reliability of data increases with an increase in the size of the data pool/set.
  • To draw reliable conclusions, multiple trials/replicates should be performed.
What students do…
  • Students take observation and develop hypothesis and an experiment design.
  • Students use scientific thinking to consider how the environment impacts gene expression and cellular networks.
  • Students conduct a controlled investigation and analyze lab results to determine how the amount of light affects Halobacterium.
  • Students compare, contrast and critique divergent results from their investigations and discuss/identify possible errors/sources of variation.  
 

Instructional Activities: (Three 50-min periods, with AT LEAST 72 hours after the 3rd period for incubation)
 
Before beginning lesson: Read entire lesson. The following documents also may help with prepping for this lab:
Advanced Prep Document – includes timing of lessons and general equipment list/suggestions
Testing light on halo-TEACHER - for help with lab set up and advance preparation if students will only be testing light/dark
Teacher Lab Hints-variables – for help with lab set-up and advance preparation if students will be selecting variables
 
Students reviewed literature about a model organism (Halo) during Lesson 1 which gave the students an opportunity to investigate the environmental conditions Halo typically grow in. The Lesson 2 laboratory experience will give students an opportunity to actually see the effect of certain environmental variables on the Halobacterium organism. Depending on the time constraints and goals for this lesson – there are 3 possible pathways. Here are the possibilities:
 
1.    Students work with a defined manipulated variable – presence of light – AND a defined, predetermined lab protocol. NOTE - This will remove one 50 minute period from the time required for this lesson, however it will be harder to present as inquiry:  observations lead to hypotheses and experimental design.
 
2.    Students work with a defined manipulated variable – presence of light – they will design a laboratory experiment using this manipulated variable which then can be carried out in the laboratory. Teacher will need to guide students with the availability of their laboratory supplies (use of stir stations, agitating incubator, bubbler system).
 
3.    Students work with different manipulated variables that are determined during their Lesson 1 research. The possible variables that can be easily manipulated using the kit (provided you have enough lab equipment) are: light, oxygen, salinity, color of light, pH, temperature, amino acid concentration, presence of metals or oil.
·         There is a document which will help outline expected results and other helpful hints for this version of the lab (Teacher Lab Hints-variables).
 
RESEARCH REFLECTION AND QUESTION FORMATION
1.    Research Reflection:
Students should refer to their notes from their Halobacterium research.  Students should be put in lab groups (3-5 students per group). They should then spend no more than 10 minutes comparing their research results to be sure the entire group has everyone’s information. This will also allow them to see that different sources may have slightly different information (e.g. ideal salinity or temperature). Students may have some holes in their research that group members can fill – any new information should be recorded. 
 
2.    Variable Selection:
After the information share, groups should come up with a list of possible, valid independent/manipulated variables that could be tested in the school’s laboratory (at least 4). Student groups should also come up with possible responding/dependent variables (at least 2). 
·         This can be done by giving student groups whiteboards or a planning format to help them get organized.
·         Remind students that only variables that make sense should be used (i.e. no aspirin, soda, etc.).
·         If time - students should brainstorm lab options (i.e. equipment needed or how would the variables be modified or measured).
 
3.      Class Discussion of Research:
This discussion should end when all important/relevant information has been gathered from the students. Students should also be sure this information is recorded. This may include the following:
·         Environmental Ideals – Temp (~37C to 42C – 37C is suggested), pH (neutral), Salinity (3-4M)
·         Ecology of Halo – Definition of an Extremophile - Extremophiles are organisms that live in harsh environments and are members of the Archaea domain. Note: this domain is not part of the true bacteria (eubacteria) domain but are made of prokaryotic cells (like eubacteria)
·         bR causes the color of Halo (this is a purple pigment). It is involved in the energy pathway that uses light.
·         Methods for creating usable energy – 3 different pathways – use of light (phototropic); use of oxygen and organic molecules (cellular respiration); anaerobic metabolism of arginine (fermentation)
 
4.      Determine possible variables for Experimental Design:
Lead students through discussion on which variables could be tested with Halobacterium.  NOTE: Depending on your plan for the lab, this can be limited to what they can do in your schools laboratory or wide open. If you allow any valid variable to be discussed, you could continue the discussion on HOW each of these would be tested (which may eliminate some choices from your list). Here are a few of the possible independent/manipulated variables you may get:
·         light, oxygen, salinity, color of light, pH, temperature, amino acid concentration, presence of pollutants (like metals or oil), carbon dioxide (this last one can lead to a conversation on whether we think this could have an effect – since they don’t use the CO2 for energy although it can have an effect on pH of the system).
The class will also need to talk about what dependant/responding variables could be measured. Possible list:  population (growth rate, amount of growth, size of population, survival rate, density, pattern of growth), color, organism size, behavior.  Consider constraints due to lab equipment available.
 
5.    Experimental Design – Using Halo to test how the environment can change population size and gene expression:
This step is determined by which path you plan to take for the lab. 
·         NOTE: If your students have no experience with the spectrophotometer – you may want to complete Lesson 5 in the Ecological Networks module to assist with the workings of the spec.
·         If your students are designing their experiment, teachers can use the following prompts to lead students to valid design.
o    Will this give you accurate results? - Students should always run three sample flasks or tubes, plus a control to be a statistically viable test.
o    Will your Halo be able to survive those conditions? Is it useful if your Halo all die in the experimental groups?
o    Why did you pick those levels of your independent/manipulated variable?
o    What is your control group?
o    Does that variable have any other impact on the environment? - e.g. temp can change pH
1.    Students working with light AND defined lab protocol:
Once students have come up with a list of variables, teacher then picks light as the manipulated variable for the class.
·         Remind students that a goal of this module is to figure out why and how the environment impacts gene expression. Tell students that while each group has a feasible experiment, the light variable is not only quite easy to control in a basic lab, but also, the gene network impacted by light and oxygen is one that we already have much data on. This means that we can combine our class data with a great deal of known data to really have a shot at answering our questions and understanding this network. Since this is our first full network analysis, it would be a good place to start and then would allow us to have more success when studying less known networks in the future.
·         The protocol for the lab can then be handed out (Testing light on Halo with stirrers-STUDENT, Testing light on Halo with incubators_STUDENT). The students can record their data in their lab notebooks using this format: Testing light on halo-STUDENT DATA SHEET or STUDENT RAW DATA SHEET. There is also a document that describes what you need to know for this protocol (Testing light on halo-TEACHER).
·         If you do not have stir stations, shaking incubator or like device, you could adapt this protocol as needed to test just light and/or oxygen also. (Intro document: Aerator Experiment Intro with Pre- and Post-lab questions, Lab investigation: Aerobic Anaerobic Light Dark Experiment with Aerator)
·         Keep in mind that when testing aerobic and anaerobic samples, your “anaerobic” sample is not truly anaerobic unless you have flushed all of the oxygen out with some suitable gas. Even though most schools do not have this capability, the sample is nearly anaerobic since, oxygen does not dissolve readily in the high salinity media for Halo. Typically, only the cells growing on the very top layer of the media in the flask have access to oxygen. Given the overall volume of the sample, this should give clear results but would be a good thing to discuss with students.
2.    Students working with light but designing lab protocol:
Once students have come up with a list of variables, teacher than picks light as the manipulated variable for the class (see note above in maroon). Students will now need to design how they will modify the light in the laboratory section. During this section students can work to design in their lab groups. Teacher will need to show students the supplies available for them to use during the experiment. In the end, teacher should help guide students to a valid protocol. This may take some time depending on how much experience your students have with designing labs. Students will most likely need additional information from you to be successful in this effort. This document (Student Lab Hints) was created as a background protocol handout to give to students. You should decide when would be the best time to give it to your students in the design process.
3.    Students working with variety of variables:
Once students have come up with a list of variables, teacher will need to help students eliminate those variables that cannot be testing with their laboratory supplies. Once the list is finalized, each group needs to select a variable from that list. The lab groups should then begin to complete their lab experimental design. Each groups should receive Student Lab Hints which will give the students information/hints for their protocol design. 
·         This process is probably best to be done initially on a white board (which will allow the teacher an easier method for approval). 
Once students have a valid experimental protocol – they can complete a pre-lab  with a rough outline of their procedure (see below note).
NOTE: Teacher’s access to equipment may limit the student’s flexibility with manipulated/independent variable levels (e.g. the kit comes with predefined salinity levels). As long as students recognize some of their steps may slightly change, this should be fine. Teacher can also describe how this models real scientific lab work where everyone must be aware of their lab’s limitations.
 
6.    Complete Lab - Using Halo to test how the environment can change population size and gene expression:
Students should now complete a lab according to the protocol they have been given or designed. Please be sure to review the document of laboratory hints/suggestions (Teacher Lab Hints-variables). Even if you are performing the defined protocol, this document may have information to assist in the success of your students’ experiments.
Due to incubation requirements, final data will be taken at the beginning of Lesson 4.  If you have equipment limitations which will only allow 1 or 2 groups to work at a time, a different timing schedule will need to be used. See the Advanced Prep document (at the beginning of module) for suggestions.
 

Students' ability to create a list of valid experimental variables based on the discussion and background research
Students' ability to create a valid experimental design for their variable
 

 
For Classrooms with an Agitating Incubator (aka Environmental Shaker/Incubator)
 
 
 

Accommodations:  For students needing help with organization, plan to use the 'team planning' worksheet.  At the end of the student lab protocol are pictures of lab equipment.  Ask students to identify when and where each piece is to be used.  The 'student lab hints' paper is one page and may be overwhelming.  Paring down may be advisable.
 
 
 

Extension Activities:  To review experimental design and thinking about variables, ask students to answer a question on plant mass.  Hand out the  question, collect answers and tally on the board, read aloud, or have students answer anonymously and then crumple their paper and toss to the other side of room. 

Students pick up and read another student's answer.  *Group students (human histogram) according to answer.  Likely answers are: light, water, air, soil, fertilizer.   To investigate the answer, use a projector to display and explore the following web-page: 

http://www.csun.edu/scied/2-longitudinal/plant_mass/index.htm#PartOne  (acknowledgement: Laurie Aiello and Brandon Duran)

   *If you allow students to text, use 'polleverywhere.com'.   Quick, free and useful for formative assessments. (Quick look)

 

 

 

 

Lesson 1 - Scientists Prepare and Plan

Posted in

Unit:  Genetics and Heredity

Part:  Environmental Influence on Gene Networks


Lesson Title: Lesson 1- Scientists Prepare and Plan


Objectives:

What students learn…

  • Observations lead to hypotheses and experimental design.
  • Halobacterium (Halo) respond to changes in their environment, such as the amount of light.

What students do…

Students use scientific thinking to consider how the environment impacts gene expression and cellular networks.

Students conduct research into current Halobacterium observations and make a list of possible variables (which leads to lesson 2).


Instructional Activities: (One 50-min period, with a bit of homework if needed)

Before beginning lesson: see Advanced Prep for help with lab set up and advance preparation for this and other lessons.

Introduction for teachers to provide context and purpose:

Students should complete these lessons near the end of a genetics unit.  Typically students would have learned that organisms (and individual cells of multicellular organisms) respond to their environment by changing which proteins they make.  A major way they do this is by regulating ”gene expression” through control of transcription.  Some genes are “turned on” to make mRNA for their corresponding protein, and some genes are “turned off” to stop making mRNA for their corresponding protein.  This allows the cell to conserve energy by producing only what is needed at that time.

This is usually studied in high school classrooms in a fairly artificial situation.  E. coli bacteria are transformed by adding plasmid DNA, and their environment is changed with the addition of the molecule arabinose.  The gene for green fluorescent protein (GFP) is “turned on” (transcribed) in the presence of arabinose and “turned off” (not transcribed) in the absence of arabinose.  This serves as a good model system to study these processes, but E. coli bacteria in nature do not have the gene for GFP, and they do not respond to the molecule arabinose in this manner.

To study this in a more natural situation where an organism’s DNA is not altered, we will use the model system of Halobacterium (Halo), which lives in high salt environments such as the Great Salt Lake.  This module will give students a way to act as scientists as they study how this organism responds to changes that occur in its natural environment.  They will use networks as part of their experimental process in the same way scientists use networks to hypothesize, model, and predict cellular responses to environmental cues.   

 UNIT DESCRIPTION AND QUESTION FORMATION

1.      Warm-up:  Pass out a pre-assessment worksheet (Pre-assessment document). Encourage students to put an answer down for each question even though they may not know the answer. (Pre-assessent Key)  Lead a class discussion based on questions they may have and specifically lead them into this question: In what ways do cells respond to their environment? Discuss also, why this is especially important for people to study today.  (Today's youth will have more information about their cells and genomes than previous generations, therefore the interplay between cells and the environment is even more important to understand and consider.)

  NOTE:  Question 1 of the pre-assessment is designed to see if students are familiar with the concept of networks.  If students need to learn about or review networks, there will be time after the lab is set up (lesson 2).  Complete the cell phone lessons 1 and 2 in Ecological Networks if needed.  Also, see lesson 3 for more information on networks.

Question 2 is one of Page Keeley assessment probes for testing whether or not students understand that systems are comprised of interactive parts.

  • Question 3 ascertains if students are familiar with the three types of data analysis involved in lesson 3.

Questions 4 and 5 are designed to see if students are familiar with experimental design.  Q4 has too many variables to be useful and in Q5 students should recognize that repeatability confirms data.

Questions 6-10 will give you an idea of how much students understand about using model organisms to better understand the interplay between the environment and the genome.

Question 10 will give you an idea of how students perceive the turning on and off of genes and what causes this.  The example matches bacterial genetics.  However, Halobacterium is not a bacteria, but an Archae and processes its genome in a similar way to eukaryotes.  For a good review of gene expression in prokaryotes and eukaryote, see this resource:  http://www.nature.com/scitable/topicpage/gene-expression-14121669.

2.  Introduction:  Power Point (Environmental Influence on Gene Networks Introduction.ppt) Use slides 2 –7 to get students thinking about the importance of gene expression and genetic control. Printable script.

Slide 2:  Define “dynamic” for students as “ever-changing.”  Allow students to offer other “whole body” changes (e.g. hair changing to grey with age, eye color changing from birth, other pre-programmed changes such as hair turning curly after puberty, or changes resulting from diseases such as the formation of cancer) and point out that if the whole body changes, and the whole body is made of cells, then cells must be changing also.

Slides 2-7:  Skin cells change by producing more melanin in response to the sun’s radiation.  Sunburn is the body’s reaction when DNA is directly damaged by UV-B light.  The damage triggers DNA repair and the production of melanin (a photo-protectant pigment) for preventing future damage.  Melanin takes UV photons and turns them into harmless amounts of heat.  The damage results in restored DNA, the replacement of new skin cells with a higher level of melanin.

Caterpillars and flamingos (shrimp) reflect color of their food source.   Both of the caterpillars are Helicoverpa armigera.  Which plant the larva feeds on influences coloration.  See 'bioone.org' research article

Flamingo feathers obtain their wonderful rosy pink color from pigments in the organisms they eat. The flamingos’ feathers, legs, and face are colored by their diet, which is rich in alpha and beta carotenoid pigments. Carotenoids in crustaceans such as those in the flamingo diet are frequently linked to protein molecules, and may be blue or green. After being digested, the carotenoid pigments dissolve in fats and are deposited in the growing feathers, becoming orange or pink. The same effect is seen when shrimp change color during cooking. The amount of pigment laid down in the feathers depends on the quantity of pigment in the flamingo’s diet. An absence of carotenoids in its food will result in new feather growth that is very pale; the existing pigment is lost through molting.

San Diego Zoo Flamingos

Drosophila wings: curly is a mutant and will develop as straight wing if pupated at 16?. 

genetics.org reprint (Lenore Ward)  see pages 1-5

http://www.journalarchive.jst.go.jp/english/jnlabstract_en.php?cdjournal=ggs1921&cdvol=31&noissue=6&startpage=163

  Hydrangeas respond to pH of soil.

Flower color in H. macrophylla is dependent on cultivar and aluminum availability. Aluminum is necessary to produce the blue pigment for which bigleaf hydrangea is noted. Most garden soils have adequate aluminum, but the aluminum will not be available to the plant if the soil pH is high. For most bigleaf hydrangea cultivars, blue flowers will be produced in acidic soil (pH 5.5 and lower), whereas neutral to alkaline soils (pH 6.5 and higher) will usually produce pink flowers. Between pH 5.5 and pH 6.5, the flowers will be purple (see image at left) or a mixture of blue and pink flowers will be found on the same plant.

http://www.usna.usda.gov/Gardens/faqs/hydrangeafaq2.html

 • Twins develop different fingerprints in response to the uterine environment.  While identical twins grow in the same uterus and have identical DNA, the placement within the uterus is different which impacts the specific formation of fingerprints. 

http://www.hhmi.org/bulletin/may2007/pdf/ask_scientist.pdf

This provides key evidence of the importance of even small environmental changes having large effects on gene expression.  We can see this manifested in the organism’s phenotype.

 • Slide 8 (Flow of Information) reminds students of how DNA results in proteins.  To answer the ‘whys’ on the previous slides, ask students to consider where in the flow of information a change would have to occur in order to get a different protein.  (Teacher note:  transcription most likely answer)

 http://www.brooklyn.cuny.edu/bc/ahp/BioInfo/GP/FlowInfo.html

Slide 9 asks students to think of ways a scientist might test how the environment impacts gene expression.  Lead students through the questions.  Brief answers: 

1)    The cell must conserve as much energy (and nutrients) as possible and does this by only making the proteins it needs, when it needs them.  Said another way, backing up from protein synthesis to the gene level, it only expresses the genes it needs, when it needs them, to save energy.  Use analogies that make sense to your students, such as, if you have 4 homework assignments due this week, you will do the one tonight that is due tomorrow instead of doing them all at the same time when that is not needed.

2)    Cells are surrounded by membranes that are typically semi-permeable.  Their local environment is the solution they are in and this changes according to diffusion/osmosis, etc.  These cells are packed in an increasingly larger environment (similar to stackable wooden dolls) that is also permeable to the outside environment.  Ways our environment impacts us: through our openings (mouth, nose, eye sockets, etc.) and even through our skin if it has a cut or even if it’s not damaged, as is the case with u-v waves.  Cells are susceptible to the outside environment.

3)    Cells change in response to stimuli from the environment.  Examples of cellular responses: movement of materials through pumps, diffusion, etc. or the movement of the cell itself via cilia, a flagellum, etc. or a change in gene expression to make a new protein or to stop or change the amount of current protein synthesis.

4)    We can put cells into an environment and systematically change that environment and measure how the cell responds.  A few examples of ways to measure that change include gene expression changes via microarray, position of cells, and growth of cells or populations. 

Slides 10-12 (Model Organisms) Discuss the use of model organisms such as fruit flies or mice.  Since changing the environment on an observable level can be difficult with a complex, multi-leveled organism, we can use model organisms.  Halobacterium is, in particular, a model organism for both experts and beginning scientists due to its non-hazardous nature.

NOTE:  Fortunately for research scientists, biological processes have been found to operate exactly the same in many different organisms. The "Krebs cycle" (the process cells use to extract energy from sugars) is the same across most species. Hemoglobin (essential for blood to carry oxygen to cells) is the same across different species of vertebrates. Because biological processes operate the same in various species, including both very simple and very complex life forms, scientists can use simpler organisms for their initial studies of biological systems.

We call this sort of simpler study case a "model organism". The simplicity of a model organism allows a scientist to more easily zero in on the properties and functions of interest, without having to sort out the complexity arising from additional systems embodied in more complex organisms. For instance, scientists can study yeast cells to understand how sugars are metabolized in many species (including in humans), without having to deal with the additional complexity from other systems in complex organisms (such as contracting muscles). Moreover, small organisms (such as yeast cells) reproduce quickly, allowing biologists to study multiple strains and generations of an organism in a short time.

Model organisms are carefully selected to provide simple cases for our initial studies of biological systems. They simplify our initial research, yet still provide data-rich and flexible experimental "systems" for us to examine. They are vital to our initial biological discoveries. Research findings from model organisms must be confirmed by also studying humans. But studies on model organisms are crucial to eventually answering the central biological questions regarding human life.

Slide 13 (Exploring how Halo...) Discuss possible variables as per slide.  The picture is of South Bay Salt Pond restoration project (near San Francisco).  You can go to “Google Earth” to get a great view of the entire area.  

Teacher Note:  Halobacterium salinarum (Halo) is not a moneran (bacteria) but belongs to Domain Archaea.

http://www.ucmp.berkeley.edu/archaea/archaea.html

Here is a short video prepared by scientist, Karlyn Beer, that may be both fun and helpful for students: http://www.youtube.com/watch?v=w-wEA4DAE3g.  

3.  Give each student the “SCIENTISTS PREPARE AND PLAN: HALOBACTERIUM EXPERIMENTS AND RESEARCH worksheet.  Outline parameters for their research, such as appropriate sites and citation as they look for information on the internet.  The following is helpful information to get students started when searching the internet:

     The wild type strain that is an ideal model organism goes by several names

o   Halobacterium salinarum (name is italicized or underlined)

o   Wild type Halobacterium

o   NRC-1

    Good search phrases:  Search the various names above or also “Halobacterium natural conditions”, “Halolophiles environment” or “Halophiles natural conditions” 

    Other key words that can be used (some alone, some together):  Archaea, salt loving, model organisms, Salt Lake Ecology, Microbiology, halobacteria, high salt environmentSee the list below (in resources) for useful scientific papers and website.  While some papers may be a stretch for students to fully understand, they can be used as a way to begin viewing scientific literature and to scan for needed pieces of information.  

    As students are researching, instead of getting frustrated by not understanding all of the terms, remind them to break words apart and to associate them with known words.  Certain words should make them think of certain environmental conditions, such as

o      Photo- = light, UV radiation = light, light driven pump = a pump that works with light

So when they see “phototaxis”, or “photosynthesis” in reference to Halo, they might consider the idea of incorporating light and dark as an environmental condition to study.

o      Fermentation = anaerobic process; whereas “oxidative” (or the like) probably involves oxygen

o      Chemo = chemicals or food in this case, so chemotaxis involves moving towards food

    If students consider the natural environment, they’ll probably come up with pollution as an environmental factor that can change gene expression.  If that is the case, steer them towards metal pollution which is common – you could experiment with nickel and iron easily

o      Nickel pollution comes from: Diesel fuel and gasoline (exhaust), lubricating oil, metal plating, bushing wear, brake lining wear, asphalt paving

o      Iron pollution comes from: Auto body rust, steel highway structures (guard rails, bridges, etc.), moving engine parts

o      Copper and other like metals are also possible as are nitrogen and phosphorus 


 
 

Assessment: How will I know they know……

Students should have compiled several possible variables gleaned from their research.  Background information should be written in notebooks and students should be ready to share findings (see lesson 2).  If students need help getting organized before class discussion, have them use a 'team planner' sheet when they discuss there ideas with their lab group. Team Plan Sheet

The preassessment will also inform you of their entry understanding of key concepts.   

 


Resources:

Advanced Prep document includes recipe for complete media and dilution information.

PowerPoint:  Environmental Influence on Gene Networks Introduction.ppt

PowerPoint Printable script 

Student Document:Pre-assessment

Teacher Document: Pre-assessment (KEY)

Lesson 1 Research Guidelines

Team Planning Document:  Team Plan Sheet

Brief, fun and helpful Halo video: http://www.youtube.com/watch?v=w-wEA4DAE3g.

Halobacterium Scientific Papers and Informational Websites:
http://genome.cshlp.org/content/11/10/1641.full

http://baliga.systemsbiology.net/ 

http://www.ebi.ac.uk/2can/genomes/archaea/Halobacterium.html 

http://www.nature.com/emboj/journal/v18/n5/pdf/7591548a.pdf

http://www.ucmp.berkeley.edu/archaea/archaea.html

http://halo.umbi.umd.edu/~haloed/


Accommodations:

If you do not have access to the internet or have students who are not capable of successful searching, print information to hand out.  Or print useful PDFs from the Extremophile Lesson 3 of the Ecological Networks module. 

This site has less complex, general information about archaeans and Halo.  It is 2 pages.  http://www.ucmp.berkeley.edu/archaea/archaea.html

If students need help getting organized before class discussion, have them use a 'team planner' sheet when they discuss there ideas with their lab group.

 

 

Picture/Image Acknowledgements:

www.eppo.org/QUARANTINE/insects/Helicoverpa_armigera/HELIAR_images.htm

http://www.brooklyn.cuny.edu/bc/ahp/SDV2.html

 


Extension Activities:

"Wanted For Being A Model Organism!" Students make a 'wanted' poster of an organism used as a model system for research (modification of activity from extensions for "Ecological Networks" Module).  Students could make a 'certificate for the model organism' in a similar fashion.  Teacher Tips  

"I Can Top That!"  Students compare and contrast what it means to be extreme.

A quick exploration of model organisms.  See lesson 3 extension for an example of how mice with gene knockouts are used to study learning and memory.  (These extensions adapted materials from Dolan DNA Learning Center at Cold Spring Harbor Laboratory)

 

 

Environmental Influence on Gene Networks Module

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Course: Biology, Genetics, Biotechnology, Environmental Science


Unit: Genetics and Heredity

Part: Environmental Influence on Gene Networks

Introduction:

In this module, students complete the steps scientists take when using systems biology to investigate how organisms induce phenotypic changes in response to the environment.   Student teams apply their background knowledge of genetics and networks to experiment with a model organism. They then exchange and interpret information in order to build a possible network, and in the laboratory, test this network by altering environmental conditions.  This leads to further experimentation to verify and draw conclusions about network interactions using experimental data and a computer simulation.  Students act as scientists while planning, implementing, and evaluating an investigation in the context of a real regulatory network.

These lessons allow students to investigate the interplay between the environment and organisms authentically, instead of through extrapolating from artifically transformed E. coli bacteria.  Through experimentation and integration of different data types, students build a comprehensive model of how genetic, cellular, and molecular components (genes, proteins, metabolites) work together in organisms and how cells function as collections of networks while being impacted by their environment. 

FLOW CHART

 

 


Objectives:

What students learn…

  • Cellular networks allow cells to respond dynamically to stimuli.
  • A response to a stimulus occurs by changing the relative amount or function of individual nodes in a cellular network.
  • A stimulus affecting one node can cause a change in many other nodes because they are all interconnected in the network.
  • Scientists use computer programs and simulations to analyze networks because networks are often very complex.
  • Some nodes have bigger system-wide affect than others.
  • Understanding relative importance of nodes allows greater understanding of how to perturb and use the system.
  • Multiple data sets from various experimental techniques are needed to show network relationships.
  • Observations lead to hypotheses and experimental design.
  • One change in the environment could affect an entire cellular network due to the interdependent nature of networks.
  • Halobacterium (Halo) respond to the amount of light in their environment
  • Halo change color from pink to purple in the presence of light and from pink to tan/white in the absence of light.
  • Halo grow better in the presence of light.
  • Using a centrifuge separates a culture into a supernatant and pellet, which allows the phenotype of cells to be more easily viewed.
  • Qualitative measurement is a way of describing data with words, whereas quantitative measurement is a way of describing data with numbers.
  • The reliability of data increases with an increase in the size of the data pool/set.
  • To draw reliable conclusions, multiple trials/replicates should be performed.
  • Mutant strains, with known genotypes, are a useful tool when studying organisms.

 

 

What students do…

  • Students use scientific thinking to consider how the environment impacts gene expression and cellular networks.
  • Students conduct a controlled investigation and analyze lab results to determine how the amount of light affects Halobacterium. 
  • Students use wild type and mutant strains of Halobacterium to investigate quantitatively and qualitatively.
  • Students use a centrifuge to spin down cells as a way to determine the phenotype of cells.
  • Students analyze various data sets to determine a bacteriorhodopsin network and to determine the importance of compiling multiple sets of data. 
  • Students compare, contrast and critique divergent results from their investigations and discuss/identify possible errors/sources of variation.
  • Students use a simulation to verify and/or correct their network understanding.
  • Students plan the steps needed for scientists to determine and verify a question using systems methods.
  • Students analyze the role the environment has on gene expression. In this analysis, recognize the components, structure, and organization of systems and the interconnections within and among them.

 


Resources:

EIGN Module Advanced Prep

Bacteriorhodopsin Network Simulation

Flowchart of Ecological Network/Gene Network combination as inquiry module

Washington State Science Standards (alignment shown)

MODULE EVALUATION

Specific documents and PowerPoint presentations located within each lesson link.

 


Pacing Guide:

This unit consists of 4 lessons designed for 50-minute periods (except where indicated). 

Lesson 1- Scientists Prepare and Plan (1 period)

          Main question explored:  What do scientists need to know before starting research of environmental impact on gene regulation using a model organism?

Lesson 2- Growth and Phenotypic Response of Halobacterium in Different Environmental Conditions (3 periods, including AT LEAST 72 hours for incubation)

            Main question explored:  In what ways do cells respond to their environment?

Lesson 3- Data Analysis to Propose Network Function (2 periods)

          Main question explored:  How do cells function as networks between genes, proteins, and the environment?

Lesson 4- Analysis of Laboratory Results to Verify Network Interactions (3 Periods)

          Main question explored:  What tools and methods are available to help scientists analyze experimental results and answer complex questions?

 
Unit Assessment:

Preassessment, formative and summative assessments located within each lesson link.

 Extension Activities: "Chroma-Challenges"

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