Assay for measuring DNA cell repair potential

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 29, 4351723, 436 63, 935 34, 935 71, 935 72, 935 82, 935 83, C12Q 168, C12P 1934, C12N 1503, C12N 1507

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049753657

ABSTRACT:
A quantitative host cell reactivation assay method comprising providing a recombinant DNA plasmid containing a bacterial gene which is transiently expressed in cells to be tested for cell repair proficiency and whose respective product assay is amenable to screening; inactivating the gene so that the gene is damaged within its coding region, the inactivation being carried out directly or indirectly by a genotoxic agent, e.g. ultra-violet light, against which cell repair proficiency is to be determined; transfecting the cells to be tested with the thus inactivated gene; allowing the transfected cells to stand for a predetermined repair period; and then determining the repair efficiency of said cells by comparing the percent gene expression with the gene expression obtained when the same plasmid DNA, without inactivation is transfected into the cells. The method may be used to test repair potential in fresh human lymphocytes or in amniotic cells.

REFERENCES:
patent: 4675285 (1987-06-01), Clark et al.
Protic-Sabljic, M. and Kraemer, K. H., (1985), PNAS 82, pp. 6622-6626.
Friedberg, E., (1985), DNA Repair, (W. H. Freeman and Co., New York, NY), pp. 505-574.
Protic-Sabljic, M. and Kraemer, K. H., (1986), Carcinogenesis, 1, No. 10, pp. 1765-1770.
Boshart et al., (1985), Cell 41, pp. 521-530.

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