High level heterologous expression in E. coli using mutant forms of the lac promoter
- PMID: 2041779
- PMCID: PMC329451
- DOI: 10.1093/nar/19.9.2417
High level heterologous expression in E. coli using mutant forms of the lac promoter
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.
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