Nucleic Acids Research, 1989, Vol. 17, No. 24 10191-10202
© 1989
Articles |
Expression of tetanus toxin fragment C in E.coli: high level expression by removing rare codons
Departments of Molecular Biology Wellcome Biotech, Langley Court, Beckenham, Kent BR3 3BS, UK 1Departments of Protein Chemistry Wellcome Biotech, Langley Court, Beckenham, Kent BR3 3BS, UK
*To whom correspondence should be addressed
Received October 20, 1989. Revised November 20, 1989. Accepted November 20, 1989.
Tetanus toxin fragment C had been previously expressed in Escherichia coli at 3–4% cell protein. The codon bias for tetanus toxin in Clostridium tetani is very different from that of highly expressed homologous genes in E. coli, resulting in the presence of many rare E. coli codons in the sequence encoding fragment C. We have replaced the coding sequence by sequence optimized for codon usage in E. coli, and show that the expression of fragment C is increased. Although the level of mRNA also increased this appeared to be a secondary consequence of more efficient translation. Complete sequence replacement increased expression to approximately 11–14% cell protein but only after the promoter strength had been improved.
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