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将小鼠次黄嘌呤磷酸核糖基转移酶基因通过DNA介导转移至培养的小鼠细胞:转移基因未整合到宿主基因组中的同源位点。

DNA-mediated transfer of the mouse gene for hypoxanthine phosphoribosyltransferase into cultured mouse cells: no integration of the transferred gene at its homologous site in the host genome.

作者信息

Willecke K, Klomfass M, Schäfer R

出版信息

Mol Gen Genet. 1981;182(1):70-6. doi: 10.1007/BF00422769.

Abstract

An established Chinese hamster cell line was fused with microcells isolated from phenotypically stable transferent mouse cells which contained a mouse transgenome coding for an abnormal form of mouse hypoxanthine phosphoribosyltransferase (HPRT, EC. No. 2.4.2.8) (Willecke et al. 1979). Two hybrids were isolated which expressed the abnormal form of mouse HPRT but no mouse alpha-galactosidase (GALA, EC. No. 3.2.1.22). In one of these microcell hybrids the abnormal HPRT activity segregated under counter-selective conditions with mouse chromosome 3. No mouse chromosome or additional mouse gene marker was found in the second microcell hybrid, possibly because of breakage and/or rearrangement of the integrated transgenome during the isolation of this hybrid. We conclude from these results that the transferred mouse HPRT gene is a phenotypically stable clone is not integrated at its homologous site on the host X chromosome. Rather, the transgenome is probably integrated into mouse chromosome 3, possibly due to homologies in repeated DNA sequences which may occur in the transgenome and which are interspersed at many sites in the host genome.

摘要

将一个已建立的中国仓鼠细胞系与从表型稳定的转移性小鼠细胞中分离出的微细胞进行融合,这些小鼠细胞含有一个编码异常形式小鼠次黄嘌呤磷酸核糖转移酶(HPRT,酶编号2.4.2.8)的小鼠转基因组(Willecke等人,1979年)。分离出两个杂种细胞,它们表达异常形式的小鼠HPRT,但不表达小鼠α-半乳糖苷酶(GALA,酶编号3.2.1.22)。在其中一个微细胞杂种中,异常的HPRT活性在反选择条件下与小鼠3号染色体分离。在第二个微细胞杂种中未发现小鼠染色体或其他小鼠基因标记,这可能是因为在该杂种分离过程中整合的转基因组发生了断裂和/或重排。从这些结果我们得出结论,转移的小鼠HPRT基因是一个表型稳定的克隆,未整合到宿主X染色体的同源位点上。相反,转基因组可能整合到了小鼠3号染色体上,这可能是由于转基因组中可能存在的重复DNA序列与宿主基因组中多个位点散布的序列具有同源性。

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