Team:Jilin China/Outline

May to July, 2014

1. MlrA gene coding optimization（codon of lactic acid bacteria）
2. MlrA gene synthesis(synthesis by pieces)
3. Sequence analysis and discussion about whether it can be split
4. Synthetic primer by company (33 pieces in total)
5. Repeat PCR for many times, recovery them and then get the complete gene product
6. Sub cloning vector and sequenced
7. Try to express by E.coli and analyze the effect of this protein
8. Express by lactic acid bacteria

July to Sept, 2014

1. Discussion about many possible paths include MC-LR
2. Try cloning Pseudomonas natural promoter.(non-coding sequences in Mlr enzyme series )
3. Synthesis by pieces then got complete product.
4. The gene transformed into E.coli and verify its functions.
5. Transform this gene by many ways into lactic acid bacteria and verify the function of promoter.