Team:NYMU-Taipei/Improve
We improved the E.coli suicide mechanism which stemmed from the 2014 Peking iGEM team and transformed pigment genes (which expressed in E.coli in past igem teams) into cyanobacteria. In suicide mechanism, we constructed the parts containing both holin and endolysin, grouped it with Nrt-A (BBa_K2350021), and even verified that the suicide mechanism worked successfully in functional test. In pigment experiments, we are the first team to transform pigment gene BBa_I742157 (CrtZ) into cyanobacteria, and even verified that the engineered cyanobacteria with BBa_I742157 had better photosynthetic efficiency.
Endolysin-Holin
Our team’s project this year is to use microalgae to produce biofuel and we proposed a mechanism to reach this goal. After a few months hard work, we can conclude that this mechanism worked successfully, and yet this is not our final target. Our team’s ultimate goal in this project is to cultivate the microalgae in an open pond so that we can have a magnificent production of biofuel. This mechanism contains transformed-E.coli and others necessary bio-bricks. And concerned about the biosafety methods in an open pond, we, therefore add in a suicide mechanism.
This mechanism which contains holin and endolysin was first proposed by 2014 Peking team. They used parts BBa_K1378031(holin) and BBa_K1378032(endolysin) for the suicide mechanism but our team has done some literature review, in the end, we decided to choose other parts of holin and endolysin which labeled as BBa_K112000 and BBa_K112806. Moreover, 2014 PeKing didn’t construct the parts containing both holin and endolysin. We also failed for a couple of times but we finally constructed it and successfully displayed in E.coli (BBa_K2350020). Not only this, we even grouped endolysin-holin’s construct with Nrt-A and created a new part (BBa_K2350021). To prove that we have successfully made up an improve part, we have done some functional test by using lactose to induce this suicide mechanism and measure the OD value of the transformed E.coli. And the result is a totally big YES! We have finally made out a functional improved part.
See more detail: Nitrogen Starvation - Suicide Mechanism Functional Test
IndC
What we are doing now in our project is not a one-step plan, we want to go further a step, and that’s why we add in the idea of pigment molecules transformation. Indigoidine, a bacterial natural product which displays in bright blue color and might be a new natural blue dye uses in industry, is the first pigment we try to transform into our cyanobacteria. This pigment we used is the existed parts of iGEM: BBa_K1152008. This part is at first only can express in E.coli, and we furthermore improve it by constructing this pigment gene sequence with a backbone pPIGBACK, which can express in cyanobacteria (BBa_K2350012).
See more detail: Pigments - Indigoidine (IndC)
CrtZ
As we said before, we are going a further step, and so on, not only blue pigment, we also try to use another type of pigment to show a different coloured algae. And what we used this time is a yellow pigment, Zeaxanthin, which is also a member of carotenoid families. CrtZ, the part we used was a part released in iGEM (BBa_I742157) . Same with indigoidine, we also construct this part on our special designed backbone pPIGBACK so that it can be expressed in our microalgae and resulted in yellow microalgae.
See more detail: Pigments - Zeaxanthin (CrtZ)
