Identification of chick thymidine kinase determinant in somatic cell hybrids of chick erythrocytes and thymidine kinase-deficient mouse cells |
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Authors: | W. -C. Leung T. R. Chen Del Rose Dubbs S. Kit |
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Affiliation: | 1. Division of Biochemical Virology, Baylor College of Medicine USA;2. Graduate School of Biomedical Sciences, University of Texas, Houston, TX 77025, USA |
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Abstract: | Disc polyacrylamide gel electrophoresis (disc PAGE) analyses of chick-mouse somatic cell hybrids [LM(TK−)/CRB]isolated from fusion mixtures of chick erythrocytes and thymidine (TdR) kinase-deficient mouse [LM(TK−)]cells have demonstrated that the somatic cell hybrids contain only chick cytosol TdR kinase F and mouse mitochondrial TdR kinase A activities. Karyotypes were analysed by the method which sequentially reveals Q- and C-bands. Four hybrid clones contained the full complement of mouse chromosomes and 1 to 3 chick micro-chromosomes. Counterselection of the LM(TK−)/CRB hybrids in 5-bromodeoxyuridine (BUdR) medium resulted in the loss of chick cytosol TdR kinase F activity and at least one of the chick chromosomes, but mouse mitochondrial TdR kinase A activity was unaffected. Unlike the LM(TK−)/CRB somatic cell hybrids, the BUdR-resistant clones could not grow in HATG (hypoxanthine-aminopte-rin-thymidine-glycine) medium. The results demonstrate that: (1) the chick cytosol TdR kinase F gene is on a member of the micro-chromosomes; and (2) selection in HATG- and BUdR-containing medium involves only cytosol TdR kinase F. |
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