Contribution

Part Contributions

The Bacillus BioBrick Box designed by LMU-Munich iGEM 2012 was used extensively in our project. The box contains three B. subtilis integrative vectors targeted to the amyE, thrC, and lacA sites, as well as constitutive and inducible promoters with varying strengths. We decided to design a fourth integrative vector and an accompanying promoter as our parts contribution.

Phag

Phag is the promoter of the hag gene, which encodes the flagellin protein . It has been shown to be one of the strongest constitutive promoters found in B. subtilis. In addition, the hag region of the genome, including flgM, is very well conserved. As such, the Phag promoter offers strong and stable protein expression.
Phag is inhibited by CsrA (“Carbon storage regulator A”), which binds at two sites of the promoter (denoted BS1 and BS2). Our designed BioBrick incorporates a single base-pair mutation in both these sites, which prevents CsrA binding.
Active Phag requires binding of the SigD transcription factor at the -10 and -35 SigD sites. When flgM binds to SigD, it prevents binding to Phag and therefore inhibits protein expression. The flgM gene is disrupted when pBSChagintegrates, thus achieving constitutive expression.

pBSChag

pBSChag is a modification of pBS1C of the BioBrick Box to integrate into the flgM site. Like pBS1C, pBSChag will have an ampicillin selection marker for E. coli and chloramphenicol for B. subtilis. The “hag” in the vector name indicates that it is designed to be used with Phag as the promoter, but others also work fine. The flgM gene is an inhibitor of Phag; pBSChag eliminates this inhibition by integrating and disrupting flgM.

LatchBio Platform

Bioinformatic processes are quickly becoming a staple in standard synthetic biology and biological science laboratories, but bioinformaticians are in an overwhelmingly short supply.

This year, the McGill iGEM team partnered with San Francisco startup LatchBio to create no-code workflows for common bioinformatic processes on the LatchBio platform.

Our team developed two workflows for genomic motif analysis. More details can be found on our software page.