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Use of a codon alteration strategy in a novel approach to cloning the Mycobacterium tuberculosis diaminopimelic acid epimerase
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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
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Official URL: http://dx.doi.org/10.1111/j.1574-6968.2006.00356.x
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 > 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: |
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| 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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