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人类12号染色体短臂cDNA的分离与区域定位

Isolation and regional assignment of human chromosome 12p cDNAs.

作者信息

Baens M, Aerssens J, van Zand K, Van den Berghe H, Marynen P

机构信息

Human Genome Analysis Laboratory, University of Leuven, Belgium.

出版信息

Genomics. 1995 Sep 1;29(1):44-52. doi: 10.1006/geno.1995.1213.

Abstract

We have characterized 117 cDNAs isolated by direct cDNA selection using pools of human chromosome 12p cosmids. Sequencing revealed that 41 clones did overlap with other cDNAs. Of the remaining 76 cDNA sequences, 11 matched previously identified human chromosome 12p genes and 3 matched previously determined cDNA sequences, including the retinoblastoma binding protein 2 (RBBP2), the cyclin-dependent kinase inhibitor KIP1, and an expressed sequence tag. For each of the 76 cDNAs specific selection by a genomic cosmid clone was confirmed. STSs were developed for all cosmids, among them 3 polymorphic simple sequence repeats associated with, respectively, the TNFR related protein, CD27, and SCNN1. Regional assignment of the STSs by PCR analysis with somatic cell hybrids and fluorescence in situ hybridization showed that the majority of the loci map to chromosome 12p13, similar to the distribution of the known 12p genes. Evidence was found for the duplication on 12p of a region containing a polymorphic simple sequence repeat and sequences of two different cDNAs.

摘要

我们已经对通过使用人12号染色体p臂黏粒文库直接进行cDNA筛选分离得到的117个cDNA进行了特征分析。测序结果显示,41个克隆与其他cDNA存在重叠。在其余76个cDNA序列中,11个与先前鉴定的人12号染色体p臂基因匹配,3个与先前确定的cDNA序列匹配,包括视网膜母细胞瘤结合蛋白2(RBBP2)、细胞周期蛋白依赖性激酶抑制剂KIP1和一个表达序列标签。对于这76个cDNA中的每一个,通过基因组黏粒克隆进行的特异性筛选均得到了证实。为所有黏粒开发了STS,其中3个多态性简单序列重复分别与TNFR相关蛋白、CD27和SCNN1相关。通过体细胞杂种PCR分析和荧光原位杂交对STS进行区域定位,结果表明大多数位点定位于12号染色体p13区,与已知的12号染色体p臂基因分布相似。发现了12号染色体p臂上一个包含多态性简单序列重复和两个不同cDNA序列的区域存在重复的证据。

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