Bacterial differentiation: Its application on production of useful substances.

细菌分化：其在有用物质生产中的应用。

基本信息

批准号：
60303024
负责人：
KOBAYASHI Yasuo
金额：
$ 4.54万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Co-operative Research (A)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

项目摘要

1. Control mechanism of bacterial differentiation (Saito, Sadafe, Fujita, Ikeuchi, Kawamura) (1) Regulatory mechanism of sporulation initiation gene spoOA expression in Bacillus subtilis was studied by the use of lacZ-fusion gene and temperature-sensitive spoOA mutant. It was shown that expression of spoOA gene depends on spoOA, OB, OE, OF, and OH genes and spprulation was not initated unless the function of SpoOA protein is derepressed. (2) spoOA gene was cloned and its gene product was overproduced in Escherichia coli and purified. Purified SpoOA protein has DNA-binding activity. (3)div gene which regulates cell division, sporulation, and protein secretion was cloned. (4) Catabolite-repressible gluconate operon was cloned. The whole nucleotide sequence was determined and the mechanism of catabolite repression of B. subtilis was studied.2. Isolation of regulatory factor of gene expression (Beppu, Nakayama) (1) Streptomycin production and sporulation in Streptomyces griseus is positively controlled by A-factor. A-factor was purified and regulatory network surrounding A-factor was studied. (2) Chromosomal proteins of B. subtilis were isolated and purified and sporulation-specific chromosomal proteins were identified.3. Development of usefulvector (Kobayashi, Yamasaki, Yamane, Kudoh, Tanaka) (1) Isolation and analysis of a temperature-dependent promoter revealed that its expressio is controlled by translational-coupling. Stable promoter-probe vector for B. subtilis was constructed. (2) A commom method to amplify a useful gene in B. Subtilis chromosome was developed. (3) Secretion vector carrying -amylase signal peptide was constructed and improvement of its secretion ability was intended. (4) prtR gene which increases protease production in B. natto was identified and cloned. It was shown that PrtR protein is a positive factor which stimulates the transcription of protease gene.

1。细菌分化的控制机制（Saito，Sadafe，Fujita，Ikeuchi，Kawamura）（1）通过使用LACZ融合基因和温度敏感的Spooa突变，研究了枯草芽孢杆菌中孢子型起始基因SPOOA表达的调节机制。结果表明，除非消除Spooa蛋白的功能，否则SPOOA基因的表达取决于SpOOA，OB，OE，OH基因和OH基因以及分支。（2）克隆Spooa基因，其基因产物在大肠杆菌中过量生产并纯化。纯化的SPOOA蛋白具有DNA结合活性。（3）克隆调节细胞分裂，孢子形成和蛋白质分泌的DIV基因。（4）克隆（克隆）可分解代谢物可抑制的葡萄糖酸酯操纵子。确定了整个核苷酸序列，并研究了枯草芽孢杆菌的分解代谢物抑制的机理。2。基因表达的调节因子的分离（Beppu，Nakayama）（1）链霉菌链霉菌素的产生和孢子体的孢子形成由A因子积极控制。纯化了A因子，并研究了A因子周围的调节网络。（2）分离并纯化枯草芽孢杆菌的染色体蛋白，并鉴定出孢子体特异性染色体蛋白。3。有用向量的开发（Kobayashi，Yamasaki，Yamane，kudoh，Tanaka）（1）依赖温度依赖性启动子的分离和分析表明，其表达是由平移耦合控制的。构建了用于枯草芽孢杆菌的稳定启动子探针载体。（2）开发了一种扩增枯草芽孢杆菌染色体中有用基因的孔隙方法。（3）构建了携带 - 淀粉酶信号肽的分泌载体，并提高了其分泌能力。（4）鉴定并克隆了纳托芽孢杆菌中蛋白酶产生的PRTR基因。结果表明，PRTR蛋白是刺激蛋白酶基因转录的积极因素。