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人晶状体内在膜蛋白MP70(Cx50)基因:克隆分析与染色体定位

The human lens intrinsic membrane protein MP70 (Cx50) gene: clonal analysis and chromosome mapping.

作者信息

Church R L, Wang J H, Steele E

机构信息

Emory University School of Medicine, Department of Ophthalmology, Atlanta, GA 30322, USA.

出版信息

Curr Eye Res. 1995 Mar;14(3):215-21. doi: 10.3109/02713689509033517.

Abstract

We have isolated and characterized a human genomic clone containing the complete coding region of lens intrinsic membrane protein MP70 (Cx50). The coding region of this DNA is completely contained within one exon, as is common of all connexins investigated to date. The size of the Cx50 coding region, from the initiating ATG to the terminating TGA is 1,299 nucleotides, coding for a polypeptide of 432 amino acids and having a translated molecular weight of 48,171 daltons. This Cx50 coding region DNA was used as a probe to analyze a panel of Southern blots of human-Chinese hamster somatic cell hybrid DNAs to assign the gene coding for Cx50 to its human chromosome. Control human and Chinese hamster DNAs displayed a distinct Eco R1 restriction fragment pattern when hybridized with the human Cx50 DNA probe. When somatic cell hybrid DNAs were restricted with Eco R1 and Southern blots hybridized with the human Cx50 DNA probe, the characteristic human restriction pattern was observed only when human chromosome 1 was present in the hybrid panel. Of the other six connexin genes which have previously been assigned to a human chromosome, two of these, Cx37 and Cx40, are also found on chromosome 1.

摘要

我们已经分离并鉴定了一个包含晶状体内在膜蛋白MP70(Cx50)完整编码区的人类基因组克隆。该DNA的编码区完全包含在一个外显子内,这是迄今为止所有已研究的连接蛋白的共同特征。Cx50编码区的大小,从起始的ATG到终止的TGA为1299个核苷酸,编码一个由432个氨基酸组成的多肽,其翻译后的分子量为48171道尔顿。这个Cx50编码区DNA被用作探针,分析一组人类-中国仓鼠体细胞杂交DNA的Southern印迹,以将编码Cx50的基因定位到其人类染色体上。当用人Cx50 DNA探针杂交时,对照的人类和中国仓鼠DNA显示出独特的Eco R1限制性片段模式。当体细胞杂交DNA用Eco R1酶切,Southern印迹用人Cx50 DNA探针杂交时,只有当杂交细胞系中存在人类1号染色体时,才观察到特征性的人类限制性模式。在先前已定位到人类染色体上的其他六个连接蛋白基因中,其中两个,Cx37和Cx40,也位于1号染色体上。

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