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小鼠精子特异性乳酸脱氢酶-X基因cDNA的分子分离及序列测定

Molecular isolation and sequence determination of the cDNA for the mouse sperm-specific lactate dehydrogenase-X gene.

作者信息

Wu K C, Chan K, Lee C Y, Lau Y F

出版信息

Biochem Biophys Res Commun. 1987 Aug 14;146(3):964-70. doi: 10.1016/0006-291x(87)90741-8.

Abstract

Several cDNA clones for the mouse lactate dehydrogenase-X (LDH-X), a sperm-specific glycolytic enzyme, were isolated from mouse testicular cDNA libraries constructed in the bacteriophage vectors, lambda gt11 and gt10. The largest cDNA clone contains an insert of 1135 base pairs in length and an open reading frame that encodes a 332 amino acid polypeptide with a molecular weight of 35.89 kD. The deduced amino acid sequence of this protein is in close agreement with the published sequence of mouse LDH-X obtained by direct protein sequencing. Northern analysis of RNA isolated from different tissues detected a single size mRNA of 1.5 kilobases in mouse testis but not in brain or liver. The Ldh-x structural gene was estimated to be about 12 kb in size as demonstrated by Southern hybridization analysis of mouse genomic DNA using the full-length cDNA as a probe.

摘要

从小鼠睾丸cDNA文库中分离出了几个编码小鼠乳酸脱氢酶X(LDH-X,一种精子特异性糖酵解酶)的cDNA克隆,该文库是用噬菌体载体λgt11和gt10构建的。最大的cDNA克隆含有一个长度为1135个碱基对的插入片段和一个开放阅读框,该开放阅读框编码一个分子量为35.89 kD的332个氨基酸的多肽。该蛋白质推导的氨基酸序列与通过直接蛋白质测序获得的已发表的小鼠LDH-X序列高度一致。对从不同组织分离的RNA进行的Northern分析在小鼠睾丸中检测到一个大小为1.5千碱基的单一mRNA,而在脑或肝脏中未检测到。用全长cDNA作为探针,通过对小鼠基因组DNA进行Southern杂交分析表明,Ldh-x结构基因的大小估计约为12 kb。

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