Team:Jilin China/Outline

May, 2014

1. Data query and project theme select
2. Experiment scheme design
3. Codon optimization of MlrA gene by lactic acid bacteria codons）
4. MlrA gene sequence design and split
5. RFP-Mlr-GFP sequence design and split

June, 2014

1. MlrA gene synthesis and sequence
2. RFP-Mlr-GFP gene synthesis and sequence

July, 2014

1. MlrA gene expression and detection
2. RFP-Mlr-GFP gene expression and detection
3. Screening of microcystin LR sensitive promoter

August, 2014

1. construction of the recombinant vector pMG-mlr
2. construction of the recombinant vector pMG-mlr
3. Synthetic primer by company (33 pieces in total)
4. Repeat PCR for many times, recovery them and then get the complete gene product
5. Sub cloning vector and sequenced
6. Try to express by E.coli and analyze the effect of this protein
7. 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.