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.
A quantitative host cell reactivation assay method comprising providing a recombinant DNA plasmid containing a bacterial gene which is transiently expressed in human lymphocytes to be tested and whose product essay is amenable to screening; inactivating said gene by exposure to ultra-violet light so that the gene is damaged within its coding region; contacting the lymphocytes to be tested with the thus inactivated gene and with diethylaminoethane (DEAE)-dextran under conditions such that transfection of said lymphocytes with the thus inactivated gene is effected; allowing the transfected lymphocytes to stand for a predetermined repair period; and then determining the repair efficiency of said lymphocytes by comparing the percent gene expression with the gene expression obtained when the same plasmid DNA, without inactivation is transfected into said lymphocytes.
Claim 2
Dependent Claim
126 words
A quantitative host cell reactivation assay method comprising providing a recombinant DNA plasmid containing a bacterial gene which is transiently expressed in human lymphocytes to be tested and whose product essay is amenable to screening; inactivating said gene by exposure to ultra-violet light so that the gene is damaged within its coding region; contacting the lymphocytes to be tested with the thus inactivated gene and with diethylaminoethane (DEAE)-dextran under conditions such that transfection of said lymphocytes with the thus inactivated gene is effected; allowing the transfected lymphocytes to stand for a predetermined repair period; and then determining the repair efficiency of said lymphocytes by comparing the percent gene expression with the gene expression obtained when the same plasmid DNA, without inactivation is transfected into said lymphocytes.
Claim 3
Dependent Claim
13 words
The method of claim 1 wherein the indicator gene codes for an enzyme.
Claim 4
Dependent Claim
12 words
The method of claim 2 wherein the plasmid is a CAT plasmid.