Open AccessExpression of tetanus toxin fragment C inE.coli: high level expression by removing rare codons
Author(s) -
Andrew Makoff,
M. D. Oxer,
Michael A. Romanos,
Neil F. Fairweather,
Stuart P. Ballantine
Publication year - 1989
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/17.24.10191
Subject(s) - biology , toxin , fragment (logic) , tetanus , escherichia coli , microbiology and biotechnology , clostridium tetani , genetics , gene , virology , vaccination , computer science , programming language
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.