When we made the draft plan about our project, we noticed that NCTU_Formosa 2010’s project made use of Cry11Aa gene. And they constructed the related part BBa_K332011. In their project, they transformed the Cry11Aa gene into E. coli to kill the larvae of mosquitoes. Considering the potential safety problem, using E. coli in the natural environment is not a good idea. Therefore, we decided to use Chlamydomonas reinhardti to express toxin genes such as Cry11Aa. Firstly, we wanted to ask them to send this part to us. And we can transform it to Chlamydomonas reinhardti to test our idea. But we noticed that the codon bias of Chlamydomonas reinhardti was significantly different from that of E. coli. At last, we decided to synthesize the codon-optimized DNA sequence to replace it. We have mentioned in the wiki that we co-expressed Cry and Cyt by 2A-peptide system. So we submit the codon-optimized Cry11Aa with 2A peptide part BBa_K2074023. Users can link the other genes by infusion technology. Based on the result of SWISS-Model, 2A peptide residues will mot affect the function of toxins.
If you are interested in the details, you can visit links as following:
We also met the same problem when we wanted to use GFP in Chlamydomonas reinhardti. To solve the low expression problem, we synthesize the eGFP based on Chlamydomonas reinhardti codon bias. We standardized it and constructed BBa_K2074026 part. If you are interested in the details, you can visit this link: http://parts.igem.org/Part:BBa_K2074026