Analysis of DNA relaxation and cleavage activities of recombinant Mycobacterium tuberculosis DNA topoisomerase I from a new expression and purification protocol

Table 2 Single stranded DNA substrates used

Substrate	Source	Primers	Substrate size (bases)	%GC
PBAD	pBAD/thio	Left-ATGCCATAGCATTTTTATCG Right-*GACCGGTACGCGTAGAATCG	556	47
MdnaK	M. tuberculosis dnaK	Left-GAACCCGTTGTTCTTAGACGAG Right-GGGTAACATCAAGCAGCAGAAC	317	62
Mtop	M. tuberculosis topA	Left-*GTAGAAGTTGTTGAGCCAGT Right-TACTCGTCGATCATCAAGAC	216	61

* Primer ³²P labeled at 5'end to analyze extended strand as cleavage substrate after denaturation of the PCR product. The left and right primers of MdnaK were utilized in separate experiments.

ISSN: 1471-2091