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Nucleic Acids Res. 1991 May 11;19(9):2417-21.

High level heterologous expression in E. coli using mutant forms of the lac promoter.

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1
Wellcome Research Laboratories, Beckenham, Kent, UK.

Abstract

Single base deletions in the lac promoter which reduced the 18bp spacing between the -35 and -10 homology regions to 17bp, increased the strength of the promoter. A single base substitution (T----G) in the -35 region to generate the consensus sequence TTG-ACA increased the strength further and no longer required a 17bp spacing. The mutated lac promoter was as powerful as a shorter form of the tac promoter which lacked two AT-rich regions upstream of the -35 region, and expressed the P69 surface antigen (pertactin) of Bordetella pertussis to 30-40% total cell protein and tetanus toxin fragment C to 16-20% total cell protein.

PMID:
2041779
PMCID:
PMC329451
[Indexed for MEDLINE]
Free PMC Article

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