We have made several contributions in this year’s project for future iGEM teams.

We constructed PBI121-COMT over-expression vector containing 35s promoter, RBS, PtoCOMT and GFP green fluorescent tags.

We also explored a method of protein-directed modification through bio-informatics and artificial intelligence.

-Application of bioinformatics in gene proofreading

After our first experiment, we discovered that our first method to transfer more COMT gene into poplar tree does not increase the efficiency of producing melatonin. We did not have time to make modification on our targeted gene, so we compute the gene sequence on multiple softwares to proofread the sequences. Then we modified some locus to produce peptide chain with greater affinity with the substrate N-acetyl serotonin than the natural enzyme. It can increase the efficiency to generate more melatonin from the poplar tree. The use of bioinformatics in gene modifying is effective because we are able to change the gene and simulate its result using software, which gives us a clear idea on our biosynthetic model.

-Leaf infiltration Method

Our goal is to increase the production of melatonin in poplar tree. The method we applied is leaf infiltration, or agrobacterium-mediated transformation. This method was thought to be deficient, since the success rate is not high enough, especially for wooden plants like poplar tree. In the experiment, our advisors and team members found out that the key is to know the cell cycle pattern of receptor material. With this pattern, agrobacterium transformation is more likely to be successful. In our experiment, the infection is successful in transporting gene into the plant. However, the efficiency of melatonin production does not increase as much as we expect. Agrobacterium-mediated transformation for wooden plant is yet to be investigated.

Parts:

We constructed a new biological compartment, which is the plasmid pBI121 attached with the COMT gene from the poplar tree.

Below are new basic parts and new composite parts as contribution to future iGEM team.

BBa_K4402000

Parts name: PtoCOMT. This is a basic part. It is a coding type.

BBa_K4402001

Parts name: RBS. This is a basic part. It is a RBS type.

BBa_K4402003

Parts name: PBI121-COMT. This is a composite part. It is a plasmid type.