1.Parts improvement
Based on the BBa_I15008 and BBa_I15009, we adjusted part of the sequence of ho1 and pcyA according to the literature and fused them for expression, which can improve the expression stability and expression of the two genes compared with the previous part BBa_M30109.Besides, an improvement was made on the basis of BBa_M30109, the excess RBS region was removed and the strongly constitutive promoter P43 (BBa_K4354003) specific to
ho1
and
pcyA
according to the literature and fused them for expression, which can improve the expression stability and expression of the two genes compared with the previous part BBa_M30109.Besides, an improvement was made on the basis of BBa_M30109, the excess RBS region was removed and the strongly constitutive promoter P43 (BBa_K4354003) specific to
Bacillus subtilis
was added, as well as our target gene NucA (BBa_K1159105) used to form the mechanism of suicide.
2.Hardware
According to the needs of our project, we have designed a hardware for sewage treatment bacteria preparation for light regulation, including sensor system, monitoring system, data display system and optical system, etc, which make it more suitable for the iteration and testing of synthetic biology, especially optogenetics. At the same time, we can upload the monitored data to the cloud platform for other users and later iGEM teams reference and use.
Graph1:3D modeling in Solidworks
Graph2:relevant data on Cloud platforms
3.Troubleshooting of sewage treatment safety
In order to solve the problems related to biosafety, reagent safety and other aspects of the sewage treatment project, the Freshwater Restoration Alliance held a meeting on biosafety issues on September 2 2022, and the participants included CFCA_China, Nanjing_NFLS, XJTLU-CHINA, Nanjing-China, CHINA-FAFU, GXU-China and NPU-CHINA.
Graph3:Biosafety online meeting
This meeting mainly discussed the safety of our project in three directions. First and foremost, to prove that our experimental steps are in accordance with safety guidelines. Second, to prove that our engineering bacteria do not cause leaks. Finally, to ensure that our projects do not cause harm to the environment and humans.
Through the full discussion of several teams, we have summarized lots of ideas for the safety of sewage treatment projects, which can be used for reference for the future iGEM teams.
Graph4: Biosafety meetup
