Robic A, Milan D, Woloszyn N, Riquet J, Yerle M, Nagel M, Bonnet M, Pinton P, Dalens M, Gellin J
INRA, Laboratoire de Génétique Cellulaire, Tolosan, France.
Anim Genet. 1997 Apr;28(2):94-102. doi: 10.1111/j.1365-2052.1997.00084.x.
Thirty-three microsatellites have been mapped on the PiGMaP porcine genetic map. By comparison with the previously published PiGMaP maps, the maps of chromosome 2 (140 cM/70 cM) and chromosome 3 (180 cM/110 cM) were extended and new markers were mapped on the p-arm extremity of chromosome 7 and on the centromeric extremity of chromosome 15. New orders are proposed for markers on chromosomes 3 and 17. Six microsatellites isolated from cosmids were also localized on the cytogenetic map by fluorescent in situ hybridization. We tested the subcloning ligation mixture-polymerase chain reaction (SLiM-PCR) method for isolating microsatellites from cosmids. Subcloning is more effective when the cosmid harbours several microsatellites whereas SLiM-PCR is more straightforward when the cosmid contains a single microsatellite. Fifteen anonymous microsatellites were regionally assigned by using a hybrid cell panel. For map integration, the determination of a regional assignment of anonymous microsatellites by using a hybrid cell panel offers an alternative to microsatellite isolation from cosmids and their localizations by in situ hybridization.
33个微卫星已被定位到PiGMaP猪遗传图谱上。通过与先前发表的PiGMaP图谱进行比较,2号染色体(140厘摩/70厘摩)和3号染色体(180厘摩/110厘摩)的图谱得到了扩展,并且新的标记被定位到7号染色体的p臂末端和15号染色体的着丝粒末端。对3号和17号染色体上的标记提出了新的排序。通过荧光原位杂交,从黏粒中分离出的6个微卫星也被定位到细胞遗传图谱上。我们测试了用于从黏粒中分离微卫星的亚克隆连接混合物 - 聚合酶链反应(SLiM-PCR)方法。当黏粒含有多个微卫星时,亚克隆更有效,而当黏粒只包含一个微卫星时,SLiM-PCR更直接。通过使用杂种细胞系对15个匿名微卫星进行了区域定位。为了进行图谱整合,通过使用杂种细胞系确定匿名微卫星的区域定位为从黏粒中分离微卫星并通过原位杂交对其进行定位提供了一种替代方法。