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编码牛晶状体内在膜蛋白(MP19)的cDNA的分子克隆及完整核苷酸序列

Molecular cloning and complete nucleotide sequence of the cDNA encoding a bovine lens intrinsic membrane protein (MP19).

作者信息

Gutekunst K A, Rao G N, Church R L

机构信息

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

出版信息

Curr Eye Res. 1990 Oct;9(10):955-61. doi: 10.3109/02713689009069931.

Abstract

Recently, we reported the partial characterization of bovine lens intrinsic membrane proteins having apparent SDS-PAGE derived molecular mass of 19, 21, and 23 kDa, and determined that they contained identical NH2- terminal amino acid sequences for the first 20 amino acids. From this amino acid sequence information, a mixed synthetic oligonucleotide was constructed and used to screen a calf lens lambda gt11 cDNA library in order to isolate and characterize the cDNA coding for this membrane polypeptide(s). Two separate cDNA clones were isolated and sequenced, and were found to have an identical sequence of 883 bases with an open reading frame coding for a polypeptide of 173 amino acids, having a molecular mass of 19,683 Daltons. The first 20 amino acids of the translated sequence were identical to that determined by our laboratory previously, and the last seven amino acids were identical to that recently determined by another laboratory from analysis of the extracted polypeptides, indicating that this cDNA is the authentic molecule coding for MP19.

摘要

最近,我们报道了牛晶状体内在膜蛋白的部分特征,这些蛋白在SDS-PAGE上的表观分子量为19、21和23 kDa,并确定它们在前20个氨基酸中含有相同的NH2末端氨基酸序列。根据这些氨基酸序列信息,构建了一个混合合成寡核苷酸,并用于筛选小牛晶状体λgt11 cDNA文库,以分离和鉴定编码该膜多肽的cDNA。分离并测序了两个独立的cDNA克隆,发现它们具有相同的883个碱基序列,开放阅读框编码一个173个氨基酸的多肽,分子量为19,683道尔顿。翻译序列的前20个氨基酸与我们实验室先前确定的相同,最后七个氨基酸与另一个实验室最近通过对提取的多肽分析确定的相同,表明该cDNA是编码MP19的真实分子。

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