Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli.
A strong promoter has been cloned on a series of plasmid vectors that facilitate the expression of cloned genes. This promoter, named tac [first described by DeBoer et al. (in Rodriguez, R.L. and Chamberlin, M.J. (Eds.),Promoters, Structure and Function. Praeger, New York, 1982, pp. pp. 462-481)] contains the -10 region of the lacUV5 promoter and the -35 region of the trp promoter. Our vectors contain various cloning sites followed by transcription termination signals. In addition, we describe plasmids that facilitate the conversion of the lac promoter to the stronger tac promoter. Thus, preexisting gene fusions using the lac or the lacUV5 promoter can be readily converted to tac promoter gene fusions without changing the ribosome-binding site (RBS). The tac promoter is repressed in lacIQ strains and can be induced by isopropylthio-beta-D-galactoside (IPTG). Studies of expression of the cI repressor of bacteriophage lambda show that the tac promoter is at least five times more efficient than the lacUV5 promoter. Under optimal conditions lambda repressor constitutes up to 30% of the total cellular protein.
D001426:Bacterial Proteins / Q000032:analysis; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D007763:Lac Operon; D010957:Plasmids
D001426:Bacterial Proteins; D004269:DNA, Bacterial