Development of 124 sequence-tagged sites and cytogenetic localization of 217 cosmids for human chromosome 10.
Development of 124 sequence-tagged sites and cytogenetic localization of 217 cosmids for human chromosome 10.
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开发了人类 10 号染色体的 124 个序列标记位点以及 217 个粘粒的细胞遗传学定位。
DOI：
10.1006/geno.1994.1345
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发表时间：
1994
期刊：
影响因子：
4.4
通讯作者：
Torrey,D
中科院分区：
文献类型：
--
作者：
Zheng,CJ;Ma,NS;Dorman,TE;Wang,MT;Braunschweiger,K;Soares,L;Schuster,MK;Rothschild,CB;Bowden,DW;Torrey,D
共有124个新的10号染色体特异序列标记位点(STS)来自两个来源：(1)从新文库中的匿名克隆获得的DNA序列，丰富了人类10号染色体的插入；(2)已发表的基因序列和其他已知的定位到10号染色体的位点的序列。文库是从携带人类10号染色体和Y染色体的体细胞杂交种构建的。通过荧光原位杂交技术，将217个宇宙染色体定位于人类10号染色体的区域。25个宇宙体代表了之前放置在基因图谱上的探测器。192个宇宙星代表了以前没有绘制过的新探测器。用标记的聚(DC-da)-聚(DG-DT)探针杂交鉴定带有CA重复的插入片段的Cosmis，并将其亚克隆以产生微卫星STS标记。构建了两个小的插入质粒库，第一个通过亚克隆从10号染色体丰富的λ噬菌体文库(LL10NS01)中插入的插入片段，第二个通过克隆从杂交种中扩增的Alu-Element介导的聚合酶链式反应产物。从克隆插入片段的DNA序列中产生STS。通过以下一个或两个标准将10号染色体特异的STS与Y染色体STS区分开来：(1)从另一个含有10号染色体的细胞系NA10926B的DNA组成的模板中成功地进行PCR扩增，或(2)FISH定位到源粘粒的10号染色体或用STS进行PCR筛选分离的YAC。这些文库是90个新的10号染色体特异STS的来源，其中42个含有CA重复序列。
A total of 124 new chromosome 10-specific sequence-tagged sites (STSs) were derived from two sources: (1) DNA sequences obtained from anonymous clones in new libraries enriched for human chromosome 10 inserts, and (2) published sequences of genes and other loci already known to map to chromosome 10. Libraries were constructed from a somatic cell hybrid carrying human chromosomes 10 and Y. A cosmid library was made from total DNA of the hybrid and probed with labeled total human DNA to identify clones with human DNA inserts. Two hundred seventeen cosmids were mapped to regions of human chromosome 10 by fluorescencein situhybridization. Twenty-five cosmids represent probes that have been placed on the genetic map previously. One hundred ninety-two cosmids represent new probes that have not been mapped previously. Cosmids carrying inserts with CA repeats were identified by hybridization with a labeled poly(dC-dA)-poly(dG-dT) probe and subcloned to yield microsatellite STS markers. Two small insert plasmid libraries were made, the first by subcloning inserts from a chromosome 10-enriched λ phage library (LL10NS01) and the second by cloningAluelement-mediated PCR products amplified from hybrid DNA. STSs were generated from the DNA sequences of clone inserts. Chromosome 10-specific STSs were distinguished from Y chromosome STSs by one or both of the following criteria: (1) successful PCR amplification from a template consisting of DNA from another chromosome 10-containing cell line, NA10926B, or (2) FISH localization to chromosome 10 of the source cosmid or of YACs isolated by PCR screening with the STS. These libraries were the source of 90 new chromosome 10-specific STSs, 42 of which contain CA repeats.