Properties of H^+-ATPase and cloning of its gene from ruminal bacteria

瘤胃细菌H^-ATP酶的性质及其基因克隆

• 批准号: 08660351
• 负责人: HINO Tsuneo
• 金额: $ 0.45万
• 依托单位: Meiji University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660351/
• 关键词: ruminal bacteria; H^+-ATPase gene; atp-mRNA; ルーメン微生物; 低pH耐性; H+^+-ATPase; unc遺伝子

A gene encoding H^+-ATPase of ruminal bacteria, Ruminococcus albus and Streptococcus bovis, was cloned and the entire operon (atp) was sequenced by the inverse PCR method. The atp operon of S.bovis was found to be approximately 6.5 kbp, which consisted of atp E (c-subunit), B (a), F (b), H (delta), A (alpha), G (gamma), D (beta), and C (epsilon) in this order. In the atp operon of R.albus, the order of atp E and B was inversed, and in addition assumed atp I (i) was included. The open reading frame was approximately 7 kbp.Primer extension analysis was conducted for the atp operon of S.bovis with the result indicating the presence of three different species of atp-mRNA.The proportion of each mRNA species was different in S.bovis cells grown at pH 4.7 compared to those grown at pH 7.0. This suggests that the synthesis of H^+-ATPase is regulated at the transcriptional or post-transcriptional level.

克隆的瘤胃细菌，瘤状球菌和bovis链球菌的H^+ - ATPase的基因被克隆，并通过逆PCR方法对整个操纵子（ATP）进行了测序。发现S.Bovis的ATP操纵子约为6.5 kbp，由ATP E（C-Subunit），B（A），F（B），H（Delta），A（Alpha），G（Gamma），D（BBETA）和C（Epsilon）组成。在R.Albus的ATP操纵子中，反向ATP E和B的顺序，此外，还包括ATP I（i）。开放的阅读框架约为7 kbp。对S.Bovis的ATP操纵子进行了Primer扩展分析，结果表明存在三种不同种类的ATP-MRNA。与在pH 7.0生长的pH值相比，在pH 4.7生长的S.bovis细胞中，每种mRNA物种的比例都不同。这表明H^+ATPase的合成受到转录或转录后水平的调节。