Team:Bulgaria/FutureWork

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Our team was inspired by the last 8 months spent on the workbench and we decided to further develop our project.Thanks to the business contacts we made, the new facility provided for us, the gene synthesis services from IDT and the skills that we acquired during our work, we believe that we can fulfil our goals. They can be separated into two major fields:

Lactobacillus sp.

• We plan to use two different strategies to transform Lactobacillus sp. with our protein sequences of interest. One of these is based on the pMG36e vector which has been tested previously in lactic acid bacteria. Our other option includes construction of pMC1-based non replicative vector that has a specific phage sequence allowing it to be integrated in the bacterial chromosome of Lactobacillus.

E.coli

• Currently we are working with the tac promoter under the control of a LacI operator. However, in our future work we would like to use also the PBADpromoter, controlled by L-arabinose as regulator for the expression of our proteins of interest.
• Since 3 of our constructs currently produce proteins in the cytoplasm of E. coli, we plan to target their expression also to: the outer side of the membrane, the inner side of the membrane and the periplasm.
• Tag the proteins for fast degradation and check these fast turn-over protein versions as osmotic stress protectants and cryoprotectants.
• Testing the activity of our proteins in cells that are subjected to various stress factors including real lyophilization conditions.
• Use CRISPR to integrate the working sequence in the bacterial chromosome.
• The main goal is to develop a fully operational method with an impact in the industry.

Moreover, we would like to see if the industry will be interested in our work. We plan to present our results to the companies that have been so far helpful to us and eventually to collaborate with them.

Last but not least, we want to continue with our efforts to popularize the synthetic biology among society. We intend to conduct more presentations about iGEM among students from different Bulgarian universities and high-schools.