Difference between revisions of "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 the suicide mechanism worked successfully in functional test. In pigment experiments, we are the first team to transform pigment genes BBa_K1152008 (IndC) and BBa_I742158 (CrtZ) into cyanobacteria, and even verified the engineered cyanobacteria with BBa_I742158 had better photosynthetic efficiency.

Endolysin-Holin

　　Our team’s project this year is about to use cyanobacteria to produce biofuel and we proposed a mechanism to reach this target. After a few month hard work, we can term to say this mechanism successfully work and yet, this is not our final target. Our team’s ultimate target in this project is to cultivate the cyanobacteria in an open pond so that we could 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 who in charge in this mechanism have done some research, in the end, we decided to choose other parts of holin and endolysin which labeled as BBa_K112000 and BBa_K112806. Moreover, they didn’t construct the parts containing both holin and endolysin. We also failed for couple of times but we finally construct it and successfully display in E.coli (BBa_K2350020), not only this, we even grouped endolysin-holin’s construct with Nrt-A and create 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 transformed E.coli. And the result is a totally big YES! We have finally made out a functional improving part.

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 use is the existed parts of iGEM: BBa_K1152008. This part is 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).

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 colour’s algae. And what we use this time is a yellow pigment, crtZ, which is also a member of carotenoid families to transform into our microalgae. The parts we use was a part released in iGEM (BBa_I742158) . Same with indigoidine, we also construct this part on our special design backbone pPIGBACK so that it can express in our microalgae and result in yellow microalgae.