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利用聚合酶链反应分析的微卫星进行大鼠基因定位。

Rat gene mapping using PCR-analyzed microsatellites.

作者信息

Serikawa T, Kuramoto T, Hilbert P, Mori M, Yamada J, Dubay C J, Lindpainter K, Ganten D, Guénet J L, Lathrop G M

机构信息

Institute of Laboratory Animals, Faculty of Medicine, Kyoto University, Japan.

出版信息

Genetics. 1992 Jul;131(3):701-21. doi: 10.1093/genetics/131.3.701.

Abstract

One hundred and seventy-four rat loci which contain short tandem repeat sequences were extracted from the GenBank or EMBL data bases and used to define primers for amplification by the polymerase chain reaction (PCR) of the microsatellite regions, creating PCR-formatted sequence-tagged microsatellite sites (STMSs). One hundred and thirty-four STMSs for 118 loci, including 6 randomly cloned STMSs, were characterized: (i) PCR-analyzed loci were assigned to specific chromosomes using a panel of rat x mouse somatic cell hybrid clones. (ii) Length variation of the STMSs among 8 inbred rat strains could be visualized at 85 of 107 loci examined (79.4%). (iii) A genetic map, integrating biochemical, coat color, mutant and restriction fragment length polymorphism loci, was constructed based on the segregation of 125 polymorphic markers in seven rat backcrosses and in two F2 crosses. Twenty four linkage groups were identified, all of which were assigned to a defined chromosome. As a reflection of the bias for coding sequences in the public data bases, the STMSs described herein are often associated with genes. Hence, the genetic map we report coincides with a gene map. The corresponding map locations of the homologous mouse and human genes are also listed for comparative mapping purposes.

摘要

从GenBank或EMBL数据库中提取了174个含有短串联重复序列的大鼠基因座,并用于设计引物,通过聚合酶链反应(PCR)扩增微卫星区域,从而创建了PCR格式的序列标签微卫星位点(STMS)。对118个基因座的134个STMS进行了特征分析,其中包括6个随机克隆的STMS:(i)使用一组大鼠×小鼠体细胞杂交克隆,将经PCR分析的基因座定位到特定染色体上。(ii)在所检测的107个基因座中的85个(79.4%)上,可以观察到8个近交大鼠品系间STMS的长度变异。(iii)基于7个大鼠回交和2个F2杂交中125个多态性标记的分离情况,构建了一个整合了生化、毛色、突变和限制性片段长度多态性基因座的遗传图谱。确定了24个连锁群,所有这些连锁群都被定位到特定的染色体上。由于公共数据库中编码序列存在偏差,本文所述的STMS通常与基因相关。因此,我们报告的遗传图谱与基因图谱相吻合。还列出了同源小鼠和人类基因的相应图谱位置,用于比较作图。

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