Use of a codon alteration strategy in a novel approach to cloning the Mycobacterium tuberculosis diaminopimelic acid epimerase

Veeraraghavan Usha, Lynn G. Dover, David L. Roper, Adrian J. Lloyd, Gurdyal S. Besra*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

Previous attempts to express the diaminopimelate epimerase gene dapF of Mycobacterium tuberculosis in Escherichia coli resulted in undetectable enzyme yields. We used silent mutation of the first 10 codons of the recombinant ORF in an attempt to reduce the formation of secondary structures that might occur near the 5′ end of the mRNA and inhibit translation. This significantly increased the yield of the enzyme, which was purified and characterized biochemically. This strategy could be generally applied to other mycobacterial genes that are difficult to express hetero-specifically and here provided pure M. tuberculosis DapF, a good foundation for future research in antimycobacterial agents.

Original languageEnglish
Pages (from-to)39-47
Number of pages9
JournalFEMS Microbiology Letters
Volume262
Issue number1
DOIs
Publication statusPublished - 1 Sep 2006
Externally publishedYes

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