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Study on Inhibition of Septum Formation of Bacteria

抑制细菌隔膜形成的研究

基本信息

批准号：
02660145
负责人：
MIYAMOTO Takahisa
金额：
$ 1.22万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02660145/
关键词：
Bacillus cereus Sporulation Septum formation Temperature sensitive DNA binding protein Chromosome

项目摘要

Bacillus cereus temperature-sensitive mutant ts-4 was obtained from Bacillus cereus JCM 2152 by the treatment with nitrosoguanidine. Synchronous chromosome replication was obtained in a culture of B. cereus ts-4 growing in a rich medium at 45ﾟC. At 10, 40, and 90 min from the start of chromosomal replication, samples of cells were transferred to a sporulation medium. After cultivation at 30 ﾟC for 20 hr, the sporulation rate was measured.The capacity of B. cereus ts-4 to sporulate was highest at 40 min after chromosome replication began, and had declined at 90 min. Other cells were cultured at 30ﾟC for 120 min after the start of replication, and their chromosomes were prepared by treatment first with 10 uglml N-acetylmuramidase SG-and 2.5 mug/ml lysozyme, and later with 1% Briji 58 and 0.4% SDS. Patterns of two-dimensional electrophoresis of the chromosomal proteins from cells transferred at 10, 40, and 90 min were compared. The chromosomes of cells transferred at 40 min had greatly decreased amounts of 10 different proteins, but the amounts of three other proteins had increased. Patterns of two-dimensional electrophoresis were also compared in de novo synthesized cromosomal proteins after the start. of chromosome replication and in DNA binding proteins prepared from B. cereus DNA cellulose column. Amounts of the same 3 different proteins greatly increased in the cells transferred at 40 min in every case. These proteins seem to control the initiation of sporulation by B. cereus ts-4.

以蜡状芽孢杆菌JCM 2152为出发菌株，经亚硝基胍诱变，获得一株温度敏感突变株ts-4。在B的培养物中获得了同步染色体复制。蜡状芽孢杆菌ts-4在45 ℃的丰富培养基中生长。在染色体复制开始后10、40和90分钟，将细胞样品转移至孢子形成培养基中。在30 ℃培养20小时后，测定其产孢率。其他细胞在30 ℃下培养120 min后，首先用10 μ g/ml N-乙酰胞壁酶SG-1和2.5 μ g/ml溶菌酶处理，然后用1%Briji 58和0.4%SDS处理，制备染色体。在10，40和90分钟转移的细胞的染色体蛋白质的二维电泳模式进行了比较。在40分钟时转移的细胞的染色体中，10种不同蛋白质的量大大减少，但另外三种蛋白质的量增加。模式的二维电泳也进行了比较，从头合成的染色体蛋白开始后。染色体复制和从B制备的DNA结合蛋白。蜡状DNA纤维素柱。在每种情况下，在40分钟转移的细胞中，相同的3种不同蛋白质的量大大增加。这些蛋白质似乎控制由B引发孢子形成。cereus ts-4。