Usha, Veeraraghavan, Dover, Lynn G., Roper, David I., Lloyd, Adrian J. and Besra, Gurdyal S. (2006) Use of a codon alteration strategy in a novel approach to cloning the Mycobacterium tuberculosis diaminopimelic acid epimerase. FEMS Microbiology Letters, Volume 262 (Number 1). pp. 39-47. doi:10.1111/j.1574-6968.2006.00356.x ISSN 0378-1097.

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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.

Item Type:	Journal Article
Subjects:	Q Science > QR Microbiology
Divisions:	Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Biological Sciences ( -2010)
Journal or Publication Title:	FEMS Microbiology Letters
Publisher:	Wiley-Blackwell Publishing Ltd.
ISSN:	0378-1097
Official Date:	September 2006
Dates:	Date Event September 2006 Published
Volume:	Volume 262
Number:	Number 1
Number of Pages:	9
Page Range:	pp. 39-47
DOI:	10.1111/j.1574-6968.2006.00356.x
Status:	Peer Reviewed
Publication Status:	Published

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