Team:USP-EEL-Brazil/Parts
We have submitted 2 new parts to the Registry. Unfortunately, due to the COVID-19 pandemic, it was not possible to perform experiments to characterize our parts.
BBa_K4091024 is a myotoxic PLA2 from Bothrops jararaca, and we intend to use it to better understand the action of this important component of snake venom.
BBa_K4091069 is a phospholipaseA2 inhibitor, and we intend to apply it to reduce the myotoxic actions of snake PLA2 in order to aid purification we added a His-TAG and, for cleavage, a TEV-site. We believe that with the advancement of research involving snake venom these new parts may be useful for future teams.
We believe that with the advancement of research involving snake venom these new parts may be useful for future teams.
We searched the sequences in the NCBI database, and identified γPLI and PLA2 from Bothrops jararaca. Since we identified the cDNA sequence it was not necessary to analyze the presence of introns. Our proteins are from snakes, eukaryotic organisms and we plan to use E. coli as chassis for expression of our proteins, for verification and removal of possible signal peptides we use the SignalP 5.0 tool from DTU Bioinformatics. The translate tool from Expasy was used to ensure that our sequence would express a functional protein after checking and removing any restriction sites for EcoRI, NotI, PstI and SpeI from the gene.
We plan to use a composite that consists of a T7 promoter with RBS (BBa K525998) and a double-terminator (BBa_B0015).