Background

Lycopene is the red pigment that gives tomatoes their color. This pigment is also made by microbes. In fact, transferring a 3-enzyme pathway to E. coli can convert farnesyl diphosphate (FPP) to lycopene. The computational tools and databases presented today can also be used to enhance lycopene production in E. coli or even produce different colors, such as the beta-carotene pigment that makes carrots orange.

For the laboratory portion of this assignment, you will characterize lycopene production in the E. coli strain from Addgene with the pAC-LYC plasmid. You will also characterize beta-carotene production in E. coli from Addgene with the pAC-BETA plasmid. Note, the pAC-LYC plasmid contains three genes from Erwinia herbicola: CrtE, CrtI, and CrtB. The pAC-BETA plasmid produces beta-carotene through the addition of the Erwinia herbicola crtY gene. All plasmids include the gene for chloramphenicol resistance.

Plasmids involved in the experiment.

Assignment

Part A: Open-Ended Questions

Part B: Plasmids 101

Part C: Labwork - Bioproduction: Plasmids, Growth, Measurement

Part A: Open-Ended Questions

1. Drew Endy’s
   1. How many people should have the option of learning to read and write DNA (i.e., practice biotechnology)
   2. Pick a country outside the United States of interest to you. How does or will its bioeconomy differ culturally and practically from the US? How do or will these differences lead to differences in opportunities for specific biotechnology projects?
   3. What is bio-scarer and does bio-scarer impact conversations about GMOs?
2. Patrick Boyle’s
   1. Assume that all of the molecular biology work you'd like to do could be automated, what sort of new biological questions would you ask, or what new types of products would you make?
   2. If you could make metric tons of any protein, what would you make and what positive impact could you have?

Part B: Plasmids 101

1. Pick a plasmid from this list of highly requested plasmids and identify:

   1. the origin of replication
   2. the antibiotic resistance
   3. the gene of interest (if it has one)
   4. the promotor
   5. the terminator/polyA sequence

   Describe in a sentence what this plasmid would be used for.

Part C: Labwork - Bioproduction: Plasmids, Growth, Measurement