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猪链球菌O-乙酰丝氨酸裂解酶编码基因的克隆与鉴定

Cloning and characterization of the gene encoding O-acetylserine lyase from Streptococcus suis.

作者信息

Osaki M, Takamatsu D, Tsuji N, Sekizaki T

机构信息

National Institute of Animal Health, Kannondai 3-1-1, Tsukuba, Ibaraki 305-0856, Japan.

出版信息

Curr Microbiol. 2000 Jan;40(1):67-71. doi: 10.1007/s002849910013.

Abstract

We have cloned and sequenced a gene encoding O-acetylserine lyase from Streptococcus suis. The gene encodes a protein of 309 amino acids with a calculated molecular mass of 32,038 Da. The deduced amino acid sequence showed more extensive similarities to the CysK proteins than to the CysM proteins of other bacteria. The cloned gene was inserted into a pTrcHisB histidine hexamer expression vector. A 38-kDa fusion protein was expressed in a cysMK auxotrophic mutant of Salmonella typhimurium and complemented the auxotrophic properties of the mutant. Furthermore, the transformants could grow in minimal defined media supplemented with not only sulfide but also thiosulfate as a sole sulfur source. These data indicated that the cloned gene encodes a protein that was a functional homolog of the CysM in S. typhimurium.

摘要

我们已经克隆并测序了猪链球菌中编码O-乙酰丝氨酸裂解酶的基因。该基因编码一个由309个氨基酸组成的蛋白质,计算分子量为32,038道尔顿。推导的氨基酸序列与其他细菌的CysK蛋白的相似性比与CysM蛋白的相似性更高。将克隆的基因插入到pTrcHisB组氨酸六聚体表达载体中。在鼠伤寒沙门氏菌的cysMK营养缺陷型突变体中表达了一种38 kDa的融合蛋白,并补充了该突变体的营养缺陷特性。此外,转化体不仅可以在补充有硫化物的基本限定培养基中生长,而且还可以在补充有硫代硫酸盐作为唯一硫源的培养基中生长。这些数据表明,克隆的基因编码一种在鼠伤寒沙门氏菌中与CysM功能同源的蛋白质。

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