Transgenic chinese hamster V79 cell lines which exhibit variable levels of gpt mutagenesis

Environmental and Molecular Mutagenesis - Tập 16 Số 1 - Trang 1-12 - 1990
Catherine B. Klein1, Toby G. Rossman1, Thomas R. Skopek1
1Institute of Environmental Medicine, New York University Medical Center, New York, New York

Tóm tắt

AbstractThe Escherichia coli gpt gene coding for xanthine‐guanine phosphoribosyl transferase has been stably transfected into HPRT− Chinese hamster V79 cells. Several gpt− cell lines have been established, which retain the sequence(s) even after long‐term culture without selection for gpt. Each cell line exhibits a characteristic spontaneous mutation frequency (10−5 to 10−2 in 6‐thioguanine (6TG) selection. While spontaneous mutagenesis to gpt− occurs rather frequently for most cell lines, it cannot be correlated with either the number of plasmid integration sites or deletion of the plasmid sequence(s). One transgenic cell line (g12), which continuously maintains a low spontaneous mutation frequency (˜3 × 10−5), was used in comparative mutagenesis studies with wild‐type V79 cells (gpt vs. hprt).Aklylating agents such as N‐methyl‐N×‐nitro‐N‐nitrosoguanidine (MNNG) and β‐propiolactone (BPL) are shown to be equally toxic and mutagenic in both g12 and V79 cells. UV and X‐rays are also equally toxic to both cell lines. The gpt locus of the g12 transfectants, however, is two to three times more sensitive to UV and 2.5—4.5 times more sensitive to X‐ray mutagenesis than the endogenous hprt of wild‐type V79 cells. The data presented here suggests that g12 cells may be useful to study mammalian mutagenesis by agents which yield limited response at the hprt locus. Future studies with these transgenic cells and other transgenic lines are planned to compare the mutability and repair of the same gene (gpt) at different integration sites in mammalian cells.

Từ khóa


Tài liệu tham khảo

10.1016/0027-5107(75)90202-X

10.1128/MCB.4.11.2266

10.1073/pnas.83.10.3356

10.1007/BF01534497

10.1111/j.1749-6632.1969.tb24893.x

10.1016/0165-1110(77)90007-0

10.1016/0022-2836(87)90668-1

Chang CC, 1978, Effects of harmon and nor‐harmon on spontaneous and ultraviolet light‐induced mutagenesis in cultured Chinese hamster cells, Cancer Res, 38, 4527

10.1016/0092-8674(74)90006-3

Christie NT, 1989, Nickel and Human Health: Current Perspectives

Cleaver JE, 1977, Induction of thioguanine‐ and ouabain‐resistant mutants and single‐strand breaks in the DNA of Chinese hamster ovary cells by 3H‐thymidine, Genetics, 87, 129, 10.1093/genetics/87.1.129

10.1016/0092-8674(82)90308-7

10.1016/0165-1110(89)90006-7

10.1093/mutage/3.1.1

10.1128/MCB.7.1.379

10.1073/pnas.83.12.4379

10.1016/0092-8674(75)90019-7

10.1002/mc.2940020404

10.1128/MCB.7.4.1459

Gillen FD, 1972, 8‐Azaguanine resistance in mammalian cells, HGPRT. Genetics, 72, 239, 10.1093/genetics/72.2.239

Glickman BW, 1987, Mammalian Cell Mutagenesis, Banbury Report 28, 167

10.1073/pnas.85.1.185

10.1289/ehp.87769

Hutchinson F, 1987, Gene Transfer Vectors for Mammalian Cells, 142

10.1128/MCB.6.5.1787

Klein CB(1988): “The Genotoxicity and Mutagenic Mechanisms of β‐propiolactone in Chinese Hamster and Bacterial Cells ” PhD. Thesis. New York University June 1988.

Klein CB, 1984, Mutagenic specificity of β‐propiolactone in bacterial and Chinese hamster V79 cells, Environ Mutagen, 6, 426

Klein CB, 1988, Loss of gpt expression in transfected V79 cells does not correlate with loss of gpt sequence, Genome, 30, 142

10.1002/em.2850140304

10.1093/nar/10.21.6763

10.1016/0027-5107(85)90101-0

10.1016/0027-5107(86)90062-X

10.1126/science.6436977

Maniatis T, 1982, Molecular Cloning: A Laboratory Manual, 1

Moore MM, 1987, Mammalian Cell Mutagenesis, Banbury Report 28, 93

10.1016/0165-1161(88)90009-X

10.1073/pnas.78.4.2072

10.1128/MCB.7.4.1445

10.1128/MCB.2.6.708

10.1016/0165-1218(88)90174-7

10.1007/BF00331159

10.1073/pnas.84.2.344

10.1016/0022-2836(77)90052-3

10.1002/em.2850110111

10.1002/em.2860060108

10.1007/BF01534640

10.1126/science.2999980

10.1016/S0022-2836(75)80083-0

10.1159/000130536

10.2307/3576723

Slankowski LF, 1987, Mammalian Cell Mutagenesis, Banbury Report 28, 71

10.1016/0027-5107(86)90037-0

10.1002/em.2860070305

Stone‐Wolff DS, 1981, Effects of inhibitors of de novo protein synthesis on UV‐mutagenesis in Chinese hamster cells. Evidence against mutagenesis via inducible, error‐prone DNA repair, Mufat Res, 82, 147

10.1016/0027-5107(85)90140-X

10.1016/0027-5107(83)90003-9

10.1016/S0027-5107(78)80021-9

Tindall KR, 1983, Cellular Responses to DNA Damage, 625

10.1016/0165-1110(89)90028-6

10.1128/MCB.4.7.1411

10.1016/0027-5107(86)90036-9

10.1021/bi00430a010

10.1073/pnas.76.3.1373

Wilson WR, 1984, Comparison of the mutagenic and clastogenic activity of amascrine and other DNA‐intercalating drugs in cultured Chinese hamster cells, Cancer Res, 44, 4420

Thông tin
Thông tin xuất bản

Environmental and Molecular Mutagenesis

Tập 16 Số 1

1-12

Thông tin tác giả