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巨大芽孢杆菌spoIIA操纵子的克隆与测序

Cloning and sequencing of the Bacillus megaterium spoIIA operon.

作者信息

Tao Y P, Hudspeth D S, Vary P S

机构信息

Department of Biological Sciences, Northern Illinois University, Dekalb 60115.

出版信息

Biochimie. 1992 Jul-Aug;74(7-8):695-704. doi: 10.1016/0300-9084(92)90142-2.

Abstract

The spoIIA operon of Bacillus megaterium has been cloned and the nucleotide sequence determined. The spoIIA sequence contains three open reading frames coding for putative proteins of 116 aa, 147 aa, and 253 aa; the first and the third genes are preceded by a ribosomal binding site. The genes are in the same order as those of B subtilis and B licheniformis. The deduced amino acid sequences of these three open reading frames show 78-92% homology with SpoIIAA, SpoIIAB and SpoIIAC of B subtilis and B licheniformis. Northern hybridization revealed that B megaterium also has two spoIIA transcripts, 1.77 kb and 2.92 kb, attaining maximum expression at t1 and t3, respectively. In addition, homology to a possible penicillin binding protein gene upstream and the first part of a spoVA operon downstream has been identified on the 3.34-kb fragment. The spoIIA and the downstream spoVA promoter regions are highly conserved among these three species. Sequence analysis of the spoVA promoter revealed a region upstream to the -35 that is highly conserved across Bacillus species.

摘要

巨大芽孢杆菌的spoIIA操纵子已被克隆并测定了核苷酸序列。spoIIA序列包含三个开放阅读框,分别编码推定的116个氨基酸、147个氨基酸和253个氨基酸的蛋白质;第一个和第三个基因之前有核糖体结合位点。这些基因的排列顺序与枯草芽孢杆菌和地衣芽孢杆菌的相同。这三个开放阅读框推导的氨基酸序列与枯草芽孢杆菌和地衣芽孢杆菌的SpoIIAA、SpoIIAB和SpoIIAC显示出78 - 92%的同源性。Northern杂交表明,巨大芽孢杆菌也有两种spoIIA转录本,分别为1.77 kb和2.92 kb,分别在t1和t3达到最大表达。此外,在3.34 kb的片段上已鉴定出与上游一个可能的青霉素结合蛋白基因以及下游spoVA操纵子第一部分的同源性。spoIIA和下游spoVA启动子区域在这三个物种中高度保守。spoVA启动子的序列分析揭示了一个位于 - 35上游的区域,该区域在芽孢杆菌属物种中高度保守。

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