Team:CSIA SouthKorea

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Hello, we are CSIA_SouthKorea!

In 2010 Collegiate iGEM, Team Cambridge conducted “E. glowi” project, in which the team used bioluminescence-related genes of firefly and Vibrio fischeri to engineer E. coli strains that emit a variety of wavelengths of light. Furthermore, Team Cambridge applied various techniques, such as mutagenesis and codon optimization, in order to improve the naturally-existing parts.

The goal of our team is to further improve the system of the parts of Team Cambridge into parts that can produce even greater amount of fluorescent protein. The main mechanism that we will utilize in order to achieve our goal is increasing the ATP concentration inside the cell. It is known from previous researches that higher concentration of ATP inside the cell leads to a greater rate of recombinant protein expression; it is also known that overexpression of PCK, which is an enzyme that synthesizes ATP under glycolytic condition, results in higher amount of ATP synthesis, which causes higher concentration of ATP inside the cell. Therefore, in order to enhance fluorescent protein expression in E. coli by increasing the ATP concentration inside E. coli, our team will improve the part engineered by Team Cambridge by incorporating PCK-expressing gene inside the part. Team Cambridge’s project comprises two sub-projects: Project Firefly and Project Vibrio; our team will specifically focus on the part with Vibrio fischeri genes.

Furthermore, considering that proteorhodopsin, under conditions in which normal cellular respiration is impossible, works to convert light energy into chemical energy of ATP, our team is considering adding gene that expresses proteorhodopsin so that we can construct E. coli that can consistently produce fluorescent protein, even when put under harsh conditions.

One plausible application of this project is constructing a BioPrinter with the bioluminescent E. coli strains. If time permits, we will try to devise a way with which we can use the E. coli strains as BioPrinter material.

Reference:

[1] Kwon, Y. D., Lee, S. Y., and Kim, P., A Physiology Study of Escherichia coli Overexpressing Phosphoenolpyruvate Carboxykinase. Biosci. Biotechnol. Biochem., 72(4), 1138-1141 (2008).

[2] Kim, H. J., Kwon, Y. D., and Lee, S. Y., An Engineered Escherichia coli Having a High Intracellular Level of ATP and Enhanced Recombinant Protein Production. Appl. Microbiol. Biotechnol., 94, 1079-1086 (2012).

[3] Walter, J. M., Greenfield, D., Bustamente, C., and Liphardt, J., Light-powering Escherichia coli with Proteorhodopsin. PNAS, 104(7), 2408-2412 (2007).

File:CSIA SouthKorea team.png

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Team CSIA_SouthKorea

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Before the meeting, through e-mail exchanges and Facebook chats, we had consolidated the direction of our project into constructing bacteria equipped with proteorhodopsin. Our advisor had sent us several theses about this topic; we tried to come up with good ideas on how to apply proteorhodopsin in making the bacteria to perform certain function. We came up with some ideas, such as enhancing butanol production in bacteria or enhancing lactic acid production in Lactobacillus. Later on, our instructor introduced us with 2010 Collegiate iGEM Team Cambridge Project, and suggested to try improving the parts constructed by Team Cambridge by adding gene that expresses proteorhodopsin or by adding other gene that enhances ATP production of E. coli. He also introduced us with [http://www.instructables.com/id/DIY-BioPrinter/ DIY BioPrinter] and suggested to create similar BioPrinter that uses bioluminescence proteins expressed by engineered E. coli instead of ink.

During the meeting, we discussed about the mechanism of DIY BioPrinter that we are planning to construct. We came up with two different mechanisms:

1. Using the bioluminescence protein-expressing bacteria itself as ink

2. Using inducers, such as arabinose and IPTG, as ink and printing on agar plate on which bacterial colonies that expresses bioluminescence proteins when contacted with inducers

The iGEM kit has arrived at the lab a few days ago, so we ook a look at it. We learned how to inoculate bacteria into solid and liquid badge, using the sample E. coli (Part A and Part B) that were included in the kit.

Results/Conclusions

What did you achieve over the course of your semester?

Safety

What safety precautions did your team take? Did you take a safety training course? Were you supervised at all times in the lab?

Attributions

Who worked on what?

Human Practices

What impact does/will your project have on the public?

Fun!

What was your favorite team snack?? Have a picture of your team mascot?

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Team:CSIA SouthKorea

From 2013hs.igem.org

Revision as of 01:24, 17 March 2013

Contents

Team

Project

Notebook

Brainstorming and Planning

Results/Conclusions

Safety

Attributions

Human Practices

Fun!