枯草芽孢杆菌中编码S-腺苷甲硫氨酸合成酶的metE基因的克隆与特性分析
Cloning and characterization of the metE gene encoding S-adenosylmethionine synthetase from Bacillus subtilis.
作者信息
Yocum R R, Perkins J B, Howitt C L, Pero J
机构信息
OmniGene Bioproducts, Inc. Cambridge, Massachusetts 02138, USA.
出版信息
J Bacteriol. 1996 Aug;178(15):4604-10. doi: 10.1128/jb.178.15.4604-4610.1996.
Abstract
The metE gene, encoding S-adenosylmethionine synthetase (EC 2.5.1.6) from Bacillus subtilis, was cloned in two steps by normal and inverse PCR. The DNA sequence of the metE gene contains an open reading frame which encodes a 400-amino-acid sequence that is homologous to other known S-adenosylmethionine synthetases. The cloned gene complements the metE1 mutation and integrates at or near the chromosomal site of metE1. Expression of S-adenosylmethionine synthetase is reduced by only a factor of about 2 by exogenous methioinine. Overproduction of S-adenosylmethionine synthetase from a strong constitutive promoter leads to methionine auxotrophy in B. subtilis, suggesting that S-adenosylmethionine is a corepressor of methionine biosynthesis in B. subtilis, as others have already shown for Escherichia coli.
摘要
通过常规PCR和反向PCR分两步克隆了来自枯草芽孢杆菌的编码S-腺苷甲硫氨酸合成酶（EC 2.5.1.6）的metE基因。metE基因的DNA序列包含一个开放阅读框，该开放阅读框编码一个400个氨基酸的序列，该序列与其他已知的S-腺苷甲硫氨酸合成酶同源。克隆的基因可弥补metE1突变，并整合到metE1的染色体位点或其附近。外源性甲硫氨酸仅使S-腺苷甲硫氨酸合成酶的表达降低约2倍。从强组成型启动子过量生产S-腺苷甲硫氨酸合成酶会导致枯草芽孢杆菌出现甲硫氨酸营养缺陷，这表明S-腺苷甲硫氨酸是枯草芽孢杆菌中甲硫氨酸生物合成的共阻遏物，正如其他人已经在大肠杆菌中所证明的那样。
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