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KMID : 0352819930090020001
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Kosin Medical Journal
1993 Volume.9 No. 2 p.1 ~ p.12
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Isolation of DNA topoisomerase II from K-562 cells and identification of cleavage sites on c-myc protooncogene
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Abstract
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DNA topoisomerase II was purified by Sephacryl S-400 gel filtration, DNA cellulose chromatography and glycerol gradient cengtrifugation from human chronic myeiogenous leukemia K-562 cells. Enzyme activity was assayed by measuring the
ATP-dependent
relaxation of supercoilled pBR322
@ES The results are summarized as follows.
@EN 1. DNA topoisomerase II was purified more than 184-fold as compared to the whole cell homogenate, with 7% yield.
2. Supercoiled pBR322 DNA was completely converted to relaxed form in the concentration of 120ng of purified DNA topoisomerase II.
3. The preferred DNA topoisomerase II cleavage sites generating the most intense bands were located in the upstream of the c-myc exon 1.
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KEYWORD
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