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通过体细胞杂交进行小鼠基因连锁分析:腺嘌呤磷酸核糖转移酶定位于8号染色体,α-半乳糖苷酶定位于X染色体。

Gene linkage analysis in the mouse by somatic cell hybridization: assignment of adenine phosphoribosyltransferase to chromosome 8 and alpha-galactosidase to the X chromosome.

作者信息

Kozak C, Nichols E, Ruddle F H

出版信息

Somatic Cell Genet. 1975 Oct;1(4):371-82. doi: 10.1007/BF01538668.

Abstract

Somatic cell hybridization techniques were applied to gene linkage analysis in the laboratory mouse. Cells of an established line of Chinese hamster lung fibroblasts were fused with mouse embryo fibroblasts and with mouse peritoneal macrophages obtained from different inbred strains. From 3 hybridization experiments, 123 primary and secondary clones were isolated in HAT selective medium and 24 were back-selected in 8-azaguanine. Hybrid clones were characterized for the expression of 16 murine isozymes by starch, acrylamide, and Cellogel electrophoresis, and on the basis of segregation data, 3 syntenic associations could be made. Malate oxidoreductase decarboxylating (MOD) and mannose phosphate isomerase (MPI) segregated concordantly, confirming an established linkage relationship; adenine phosphoribosyltransferase (APRT) segregated concordantly with glutathione reductase (GR) which is known to be on chromosome 8; alpha-galactosidase was observed to be syntenic with hypoxanthine phosphoribosyltransferase (HPRT), and X-linked enzyme. All other isozymes examined segregated independently of one another.

摘要

体细胞杂交技术被应用于实验室小鼠的基因连锁分析。将已建立的中国仓鼠肺成纤维细胞系的细胞与小鼠胚胎成纤维细胞以及从不同近交系获得的小鼠腹腔巨噬细胞融合。在3次杂交实验中,在HAT选择培养基中分离出123个初级和次级克隆,并在8-氮杂鸟嘌呤中进行了24次回选。通过淀粉、丙烯酰胺和Cellogel电泳对杂交克隆进行了16种小鼠同工酶表达的表征,根据分离数据,可以建立3种同线关联。苹果酸氧化还原酶脱羧酶(MOD)和磷酸甘露糖异构酶(MPI)协同分离,证实了已建立的连锁关系;腺嘌呤磷酸核糖转移酶(APRT)与已知位于8号染色体上的谷胱甘肽还原酶(GR)协同分离;观察到α-半乳糖苷酶与次黄嘌呤磷酸核糖转移酶(HPRT,一种X连锁酶)同线。所有其他检测的同工酶彼此独立分离。

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