One step engineering of T7-expression strains for protein production: increasing the host-range of the T7-expression system.

ABSTRACT: The T7-expression system has been very useful for protein expression in Escherichia coli. However, it is often desirable to over-express proteins in species other than E. coli. Here, we constructed an inducible broad-host-range T7-expression transposon, which allows simple one-step construction of T7-expression strains in various species, providing the option to over-express proteins of interest in a broader host-range. This transposon contains the T7 RNA polymerase driven by the lacUV5 promoter, which is repressed by the lac-repressor. Leaky expression is prevented by the presence of T7-lysozyme on this construct. The complete T7-expression system is flanked by mariner transposon repeats of the suicidal R6Kgammaori plasmid, pBT20-Deltabla. Stable integration of the whole system is possibl

SUBMITTER: Kang Y 

PROVIDER: S-EPMC2040312 | biostudies-literature | 2007 Oct

REPOSITORIES: biostudies-literature