Team:Pitt/HSP60 Promoter/Intro

From 2014.igem.org

HSP60 Promoter Intro

Essential to using Propionibacterium acnes as a new chassis for synthetic biology applications was to first identify genetic regulatory elements that could lead to efficient transgene expression in P. acnes. P. acnes is very distantly related to E. coli, belonging to a different phylum. Thus, we hypothesized that E. coli promoters would not function in P. acnes. Additionally, while some P. acnes promoters have predicted in the recently sequenced genome, none have been cloned to date.(Ref 1) Therefore we sought to find promoter elements from the closely-related and well-characterized mycobacterium M. bovis. Specifically, we cloned the Hsp60 promoter from M. bovis, a promoter known to be highly active in mycobacteria and to function in other propionibacteria.(Ref 2) As we didn’t know what RBS’s would be optimal for P. acnes we generated two versions of the promoter - one which contained only the promoter sequence and no RBS (Part: BBa_K1548000), and another which contained the promoter and the strong RBS from bacteriophage TM4 (Part: BBa_K1548001), known to lead to high levels of translation in mycobacteria.