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木醋杆菌磷酸葡萄糖变位酶基因的核苷酸序列与表达分析

Nucleotide sequence and expression analysis of the Acetobacter xylinum phosphoglucomutase gene.

作者信息

Brautaset T, Standal R, Fjaervik E, Valla S

机构信息

Unigen Center for Molecular Biology, University of Trondheim, Norway.

出版信息

Microbiology (Reading). 1994 May;140 ( Pt 5):1183-8. doi: 10.1099/13500872-140-5-1183.

Abstract

The Acetobacter xylinum gene (celB) encoding phosphoglucomutase (EC 5.4.2.2) has previously been cloned by complementation of cellulose-negative mutants. In the present report the nucleotide sequence of a 2.0 kb DNA fragment containing celB is described. Expression analysis using the bacteriophage T7 RNA polymerase promoter phi 10 resulted in identification of a probable translational start codon of celB, and this conclusion was confirmed by N-terminal amino acid sequencing of the recombinant protein. From the nucleotide sequence data it was deduced that celB encodes a protein with a calculated molecular mass of 59.6 kDa. A protein of similar size was visualized after in vitro transcription and translation, using the cloned 2.0 kb fragment as template. The results of an amino acid sequence comparison and a biochemical analysis indicated that the CelB protein is structurally and functionally related to the previously characterized human and rabbit phosphoglucomutases.

摘要

先前已通过纤维素阴性突变体的互补作用克隆了编码磷酸葡萄糖变位酶(EC 5.4.2.2)的木醋杆菌基因(celB)。在本报告中,描述了包含celB的2.0 kb DNA片段的核苷酸序列。使用噬菌体T7 RNA聚合酶启动子phi 10进行的表达分析确定了celB可能的翻译起始密码子,并且通过重组蛋白的N端氨基酸测序证实了这一结论。从核苷酸序列数据推断,celB编码一种计算分子量为59.6 kDa的蛋白质。以克隆的2.0 kb片段为模板,在体外转录和翻译后观察到了大小相似的蛋白质。氨基酸序列比较和生化分析结果表明,CelB蛋白在结构和功能上与先前鉴定的人和兔磷酸葡萄糖变位酶相关。

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