编码牛丁二醇脱氢酶的cDNA的分子克隆与特性分析

Molecular cloning and characterization of a cDNA encoding a bovine butanediol dehydrogenase.

作者信息

Smania A M, Argaraña C E

机构信息

Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Argentina.

出版信息

Gene. 1997 Sep 15;197(1-2):231-8. doi: 10.1016/s0378-1119(97)00267-9.

DOI:10.1016/s0378-1119(97)00267-9

PMID:9332371

Abstract

Using a polyclonal antibody against a bovine brain 30-kDa protein (p30), we isolated from a lambda gt11 bovine brain expression library a cDNA that codifies a protein with an apparent molecular mass of 30 kDa. The cDNA nucleotide sequence contained a unique open reading frame encoding a 26.7 kDa polypeptide. The 257 amino acids deduced sequence showed a significant homology with several dehydrogenases, mainly with a bacterial acetoin reductase (62%). The cloned cDNA identity was confirmed by the determination of acetoin reductase activity in lysogens of lambda phage constructions containing the full length cDNA. The results described in this report are to our knowledge the first molecular characterization of a 2,3-butanediol dehydrogenase in mammals.

摘要

我们使用针对牛脑30 kDa蛋白（p30）的多克隆抗体，从λgt11牛脑表达文库中分离出一个cDNA，该cDNA编码一种表观分子量为30 kDa的蛋白质。该cDNA核苷酸序列包含一个独特的开放阅读框，编码一个26.7 kDa的多肽。推导的257个氨基酸序列与几种脱氢酶具有显著同源性，主要与细菌乙酰甲基甲醇还原酶同源（62%）。通过测定含有全长cDNA的λ噬菌体构建体溶原菌中的乙酰甲基甲醇还原酶活性，证实了克隆的cDNA的同一性。据我们所知，本报告中描述的结果是哺乳动物中2,3-丁二醇脱氢酶的首次分子特征描述。