通过脉冲场凝胶电泳对与囊性纤维化基因座紧密连锁的两个DNA标记进行物理定位。

Physical localization of two DNA markers closely linked to the cystic fibrosis locus by pulsed-field gel electrophoresis.

作者信息

Rommens J M, Zengerling-Lentes S, Kerem B, Melmer G, Buchwald M, Tsui L C

机构信息

Department of Genetics, Hospital for Sick Children, University of Toronto, Ontario, Canada.

出版信息

Am J Hum Genet. 1989 Dec;45(6):932-41.

Our previous linkage analysis suggested that the DNA segment D7S122 is located between MET and D7S8, the two genetic markers that are thought to flank the cystic fibrosis locus (CF). Subsequent chromosome walking experiments revealed that D7S122 in within close distance to another randomly isolated DNA marker, D7S340. To determine the physical relationship among D7S122, D7S340, MET, and D7S8, we have constructed a long-range restriction map of the region containing these four DNA segments, by using DNA from a human/hamster somatic hybrid cell line 4AF-KO15 (containing a single human chromosome 7) and a series of rare-cutting restriction enzymes. The combined results of complete, partial, and double digestion analyses confirm that D7S122 and D7S340 are located between MET and D7S8. The order of these markers is MET-D7S340-D7S122-D7S8, with distance intervals of approximately 500, 10, and 980 kbp, respectively. Together with family analysis, this information will be useful for eventual identification of the CF gene.

我们之前的连锁分析表明，DNA片段D7S122位于MET和D7S8之间，这两个遗传标记被认为位于囊性纤维化基因座（CF）两侧。随后的染色体步移实验表明，D7S122与另一个随机分离的DNA标记D7S340距离很近。为了确定D7S122、D7S340、MET和D7S8之间的物理关系，我们利用来自人/仓鼠体细胞杂种细胞系4AF-KO15（含有一条人类7号染色体）的DNA和一系列稀有切割限制酶，构建了包含这四个DNA片段区域的长程限制酶切图谱。完全酶切、部分酶切和双酶切分析的综合结果证实，D7S122和D7S340位于MET和D7S8之间。这些标记的顺序是MET-D7S340-D7S122-D7S8，距离间隔分别约为500、10和980千碱基对。结合家系分析，这些信息将有助于最终鉴定CF基因。