BioBuilding: Synthetic Biology for Students: Lab 5: Difference between revisions
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By the conclusion of this laboratory investigation, the student will be able to: | By the conclusion of this laboratory investigation, the student will be able to: | ||
* Define and properly use synthetic biology terms: chassis, system, device, redundancy | * Define and properly use synthetic biology terms: chassis, system, device, redundancy | ||
* Define and properly use molecular genetics terms: PCR, gene expression, codon | * Define and properly use molecular genetics terms: PCR, gene expression, codon shuffling. | ||
* Explain the role of redundancy in synthetic biology and engineering. | * Explain the role of redundancy in synthetic biology and engineering. | ||
* Conduct PCR and TLC and interpret the results each. | * Conduct PCR and TLC and interpret the results each. | ||
* Compare two engineering solutions to a given problem (redundancy vs kill switches) | |||
==Introduction== | ==Introduction== | ||
"Nature is a masterful and prolific chemist" (Microbiol. Mol. Biol. Rev. March 2005 vol. 69 no. 1 51-78, [doi: 10.1128/MMBR.69.1.51-78.2005] and many laboratories work hard to mimic even the smallest bit of nature's range and skill. In this experiment we'll examine the biosynthesis of carotenoids, a chemical family in the isoprenoid family that's responsible for many of the vibrant colors seen in plants and animals. Nature makes it look easy! Think of the bright orange color of carrots and you're thinking of the carotenoid they make called beta-carotene. There are more than 600 natural carotenoids, playing important roles in harvesting light for photosynthesis, as anti-oxidants to detoxify reactive species, and as regulators of membrane fluidity. The structure of carotenoids makes them lipophilic so in the lab they're more soluble in organic solvents like acetone than they are in water. We'll exploit this fact when we measure the beta-carotene in a collection of cells that we'll grow. | |||
Plants can make their own carotenoids from scratch, but animals can't so we must eat all we need. That can lead to vitamin deficiencies. In cultures that can't grow many vitamin-rich plants, individuals can develop illnesses related to vitamin deficiency. You may want to consider biotechnology approaches to this issue, including the story of "golden rice" and the social impact of GMOs in the US and in Europe. | |||
[[Image:Metabolic Pathway for b-carotene.png]] | [[Image:Metabolic Pathway for b-carotene.png]] | ||
==Procedure== | ==Procedure== | ||
====Part 1: Testing Genetic Variability==== | ====Part 1: Testing Genetic Variability==== | ||
A video showing you how to restreak cells is [http://youtu.be/bfKUUShF2-M here.] | |||
#Yeast will arrive on a YPD plate to grow at 30°C or room temp and stored at room temp or in the fridge. | #Yeast will arrive on a YPD plate to grow at 30°C or room temp and stored at room temp or in the fridge. | ||
#Identify color variants and restreak onto fresh YPD. Are there differences in the stability of the phenotypes? Are there growth conditions that make the colors more or less stable? | #Identify color variants and restreak onto fresh YPD. Are there differences in the stability of the phenotypes? Are there growth conditions that make the colors more or less stable? | ||
Revision as of 09:59, 20 February 2013
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Eau That Smell Lab |
Lab 5: Golden Bread
Acknowledgments: This lab was developed with materials from the Johns Hopkins 2011 iGEM team, as well as guidance and technical insights from BioBuilder teachers around the countryObjectivesBy the conclusion of this laboratory investigation, the student will be able to:
Introduction"Nature is a masterful and prolific chemist" (Microbiol. Mol. Biol. Rev. March 2005 vol. 69 no. 1 51-78, [doi: 10.1128/MMBR.69.1.51-78.2005] and many laboratories work hard to mimic even the smallest bit of nature's range and skill. In this experiment we'll examine the biosynthesis of carotenoids, a chemical family in the isoprenoid family that's responsible for many of the vibrant colors seen in plants and animals. Nature makes it look easy! Think of the bright orange color of carrots and you're thinking of the carotenoid they make called beta-carotene. There are more than 600 natural carotenoids, playing important roles in harvesting light for photosynthesis, as anti-oxidants to detoxify reactive species, and as regulators of membrane fluidity. The structure of carotenoids makes them lipophilic so in the lab they're more soluble in organic solvents like acetone than they are in water. We'll exploit this fact when we measure the beta-carotene in a collection of cells that we'll grow. Plants can make their own carotenoids from scratch, but animals can't so we must eat all we need. That can lead to vitamin deficiencies. In cultures that can't grow many vitamin-rich plants, individuals can develop illnesses related to vitamin deficiency. You may want to consider biotechnology approaches to this issue, including the story of "golden rice" and the social impact of GMOs in the US and in Europe.
ProcedurePart 1: Testing Genetic VariabilityA video showing you how to restreak cells is here.
Part 2: PCR
Part 3: Yeast TransformationPart 4: Measuring Vitamin APart 5: Baking BreadNext dayIn your lab notebook, you will need to construct a data table as shown below. These may be provided. Also be sure to share your data with the BioBuilder community here. Lab ReportI. Introduction
II. Methods
III. Results
IV. Discussion
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