| Publication number |
CLPUB00677 |
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| Authors |
Jha K.K., Ozer H.L. |
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| Title |
Genetic studies with a mutant mouse cell, ts-2 Balb/3T3, with a temperature sensitive defect in DNA synthesis. |
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| Citation |
Genetics 86 Suppl. 2:S32-S33(1977) |
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| Web pages |
https://eurekamag.com/research/026/681/026681029.php |
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| Abstract |
ts-2 permits cellular and polyoma viral DNA synthesis at 33 Celsius but
not at 38 Celsius. Somatic cell hybrids of ts-2 and a non ts 3T3 which is
thioguanine-resistant (Tgr) and ouabain resistant (OUAr) can be isolated
at 33 Celsius in selective medium containing hypoxanthine, aminopterin,
thymidine (HAT) plus ouabain. Such hybrids are able to grow at 38 Celsius
demonstrating the recessive nature of the ts-2 phenotype. For obtaining
hybrids between ts-2 and other ts or normal cells of rodent or human
origin the TGr (inability to grow in HAT medium) and OUAr markers were
induced in the ts-2 cell line. Somatic cell hybrids between this doubly
marked ts-2 cell line, 2TGO-13, and human fibroblasts were isolated in
HAT, ouabain (10^-5M) medium at 33 Celsius. The human ts-2 hybrid cells
were thioguanine-sensitive and back selection for thioguanine-resistance
(loss of hypoxanthine-guanine phosphoribosyltransferase gene on the human
X chromosome) resulted in loss of temperature-resistance of hybrid cells
in greater than 95% of the hybrid colonies, suggesting synteny of the two
genetic markers. Cell hybrids between ts-2 and five other mouse cells with
ts phenotype for DNA synthesis do not show complementation for the ts
defect.
|
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| Cell lines |
CVCL_B5VE; BALB/3T3 clone A31 ts2 CVCL_B5VK; BALB/3T3 clone A31 ts2-TGO |
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