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The genetic manipulation of  Mycobacterium tuberculosis  genome is limited by the availability of selection markers. Spontaneous resistance mutation rate of  M. tuberculosis  to the widely used kanamycin is relatively high which often leads to some false positive transformants. Due to the few available markers, we have created a cassette containing thiostrepton resistance gene ( tsr ) for selection in  M. tuberculosis  and  M. bovis  BCG, and gentamicin resistance gene ( aacC1 ) for  Escherichia coli  and  M. smegmatis  mc 2 155, flanked with  dif  sequences recognized by the Xer system of mycobacteria. This cassette adds to the limited available selection markers for mycobacteria.

A Cassette Containing Thiostrepton, Gentamicin Resistance Genes, and dif sequences Is Effective in Construction of Recombinant Mycobacteria