Team:BostonU/Data

We are also introducing a new promoter and gene pair to iGEM that forms a copper-dependent repressible system. Since the library of metal sensitive promoters was relatively small on the Registry of Standard Biological Parts, we decided to investigate a metal-dependent system. We chose mmoR and a σ⁵⁴ promoter associated with it from a bacterium called Methylosinus trichosporium OB3b. mmoR is a σ⁵⁴-dependent transcriptional activator.

It is unclear if the σ⁵⁴ promoter will function in E. coli in the absence of mmoR. nBLAST results comparing the mmoR to the E. coli genomes available yielded very short hits, with the top hit showing matches for less than 200bp out of the possible 2070bp length of mmoR. However, the pBLAST hits showed stronger hits with other σ⁵⁴-dependent transcriptional activators found in E. coli. The top nBLAST and pBLAST hits are shown below.