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PMID: 2690015 Published · ppublish English Journal Article

Expression of tetanus toxin fragment C in E. coli: high level expression by removing rare codons.

Nucleic acids research ·Vol. 17 ·No. 24 ·1989-12-25 ·Pages 10191-202

Makoff AJ, Oxer MD, Romanos MA, Fairweather NF, Ballantine S

Abstract

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.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular Codon Escherichia coli/genetics Gene Expression Molecular Sequence Data Nucleic Acid Hybridization Peptide Fragments/genetics Plasmids Promoter Regions, Genetic RNA, Messenger/genetics Tetanus Toxin/genetics Transcription, Genetic
Chemicals
Codon Peptide Fragments RNA, Messenger Tetanus Toxin tetanus toxin fragment C
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Makoff A J
Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, UK.
Oxer M D
Romanos M A
Fairweather N F
Ballantine S
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/17.24.10191
Published
1989-12-25
Pages
10191-202
Language
English
Region
England
NLM ID
0411011
PMCID
PMC335293
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/17.24.10191 PMC Full Text View journal details
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