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单纯疱疹胸苷激酶基因与转化人细胞中18号染色体的关联。

Association of herpes simplex thymidine kinase gene with chromosome No. 18 in transformed human cells.

作者信息

McKinlay M A, Wilson D E, Harrison B, Povey S

出版信息

J Natl Cancer Inst. 1980 Feb;64(2):241-8. doi: 10.1093/jnci/64.2.241.

Abstract

The herpes simplex virus type-2 (HSV-2)-transformed human cell line HB-2-3 was fused with thymidine kinase (TK)-deficient mouse cells [LM(TK-)], and 12 independent hybrids were isolated with the use of the HAT (hypoxanthine, amethopterin, and thymidine)-ouabain selection system. Discontinuous polyacrylamide gel electrophoresis studies demonstrated that the HSV-2-specific TK was the selected enzyme in the hybrids. Isoenzyme analysis and karyotyping were used in the analysis of the hybrids for the presence of human chromosomes. All 12 hybrids contained human chromosome No. 18 and the enzyme peptidase A, which is encoded by a gene on this chromosome. Hybrids were exposed to bromodeoxyuridine (BrdUrd) as a means of selection for cells that had lost HSV-2 TK activity. Isoenzyme and karyotyping data obtained from 33 BrdUrd-resistant sublines were consistent with the hypothesis that the HSV-2 TK gene is associated with chromosome No. 18 in the HB-2-3 cell line.

摘要

将单纯疱疹病毒2型(HSV - 2)转化的人细胞系HB - 2 - 3与缺乏胸苷激酶(TK)的小鼠细胞[LM(TK -)]进行融合,并使用HAT(次黄嘌呤、氨甲蝶呤和胸腺嘧啶核苷)-哇巴因选择系统分离出12个独立的杂种细胞。不连续聚丙烯酰胺凝胶电泳研究表明,HSV - 2特异性TK是杂种细胞中被选择的酶。在分析杂种细胞中人类染色体的存在时,使用了同工酶分析和核型分析。所有12个杂种细胞都含有18号人类染色体和肽酶A,该酶由这条染色体上的一个基因编码。将杂种细胞暴露于溴脱氧尿苷(BrdUrd)中,以此作为选择失去HSV - 2 TK活性细胞的一种方法。从33个抗BrdUrd亚系获得的同工酶和核型分析数据与以下假设一致:在HB - 2 - 3细胞系中,HSV - 2 TK基因与18号染色体相关。

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