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KMID : 0380219940270030196
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Journal of Biochemistry and Molecular Biology
1994 Volume.27 No. 3 p.196 ~ p.200
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Development of An In Vitro System for Detection of Deletions Mediated by DNA Topoisomerase II
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Abstract
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Abstract:
@EN A 23-mer DNA containing a topoisomease II consensus sequence was synthesized in order to examine whether topoisomerase II mediates deletion formation between toposiomerase II consensus sequences. Using a synthetic DNA duplex, which can be
cleaved by
calf thymus topoisomerase II. Shuttle vector pBK3-2 was developed which contains the two bacterial makers amp and neo, the replication origins of pBR322 and SV40 DNA, and topoisomerase II consensus sequence. This plasmid allowed a test of whether
topoisomerase II mediates deletion fromation between between topoisomerase II consensus sequences. In this system deletion formation was observed at a frequency of 3.5×10E4 per total Amp' colonies. This frequency is significantly higher than of
deletion formation in reaction mixtures without topoisomerase ll. Topoisomerase II inhibitor VM26 inhibited deletion formation.
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KEYWORD
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