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通过细胞融合将麂属基因片段插入仓鼠细胞。

Insertion of muntjac gene segment into hamster cells by cell fusion.

作者信息

Tsukamoto K, Klein R, Hatanaka M

出版信息

J Cell Physiol. 1980 Aug;104(2):225-32. doi: 10.1002/jcp.1041040212.

Abstract

Indian muntjac diploid cells that have only three pairs of easily discernible large chromosomes were fused with hamster cells deficient in hypoxanthine guanine phosphoribosyltransferase (HGPRT) using polyethylene glycol. Cells that survived in hypoxanthine-aminopterine-thymidine (HAT)-oubaine medium were analyzed. Hybrid cells containing both muntjac and hamster chromosomes in a given cell were not found. Instead, the cells had the same chromosomal sets as those of either parental muntjac or hamster cells. A clonal isolate that had the same chromosomal sets as those of parental hamster cells were analyzed in detail and showed the following characteristics: (1) portions of the survival curve in various concentrations of HAT medium were intermediate between those of parental cells; (2) expressions of both muntjac and hamster antigen(s) were detected by immunofluorescence staining; (3) the mobility of the enzyme HGPRT in gel electrophoresis differed from that of parental hamster or muntjac cells. These results indicate that the clonal isolate (AD202h) is a somatic cell hybrid of hamster and muntjac that contains chromosomal sets of hamster with an inserted segment of the muntjac genome, including HGPRT. The formation of such an unusual hybrid and a possible explanation of transfer of some gene segments in the hybrid cell in this system are discussed.

摘要

利用聚乙二醇将仅具有三对易于辨认的大染色体的印度麂二倍体细胞与缺乏次黄嘌呤鸟嘌呤磷酸核糖转移酶(HGPRT)的仓鼠细胞融合。对在次黄嘌呤 - 氨基蝶呤 - 胸腺嘧啶核苷(HAT) - 乌本苷培养基中存活的细胞进行了分析。未发现给定细胞中同时含有麂和仓鼠染色体的杂交细胞。相反,这些细胞具有与亲本印度麂或仓鼠细胞相同的染色体组。对一个具有与亲本仓鼠细胞相同染色体组的克隆分离株进行了详细分析,其表现出以下特征:(1)在不同浓度的HAT培养基中的存活曲线部分介于亲本细胞之间;(2)通过免疫荧光染色检测到印度麂和仓鼠抗原的表达;(3)HGPRT酶在凝胶电泳中的迁移率与亲本仓鼠或印度麂细胞不同。这些结果表明,该克隆分离株(AD202h)是仓鼠和印度麂的体细胞杂种,其包含带有插入的印度麂基因组片段(包括HGPRT)的仓鼠染色体组。讨论了这种异常杂种的形成以及该系统中杂种细胞中一些基因片段转移的可能解释。

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