Molecular cloning and sequencing of the upstream region of the major Bacillus subtilis autolysin gene: a modifier protein exhibiting sequence homology to the major autolysin and the spoIID product.
Molecular cloning and sequencing of the upstream region of the major Bacillus subtilis autolysin gene: a modifier protein exhibiting sequence homology to the major autolysin and the spoIID product.
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主要枯草芽孢杆菌自溶素基因上游区域的分子克隆和测序：一种与主要自溶素和 spoIID 产物表现出序列同源性的修饰蛋白。
DOI：
10.1099/00221287-138-6-1067
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发表时间：
1992
期刊：
影响因子：
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通讯作者：
J. Sekiguchi
中科院分区：
文献类型：
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作者：
A. Kuroda;M. H. Rashid;J. Sekiguchi
通过染色体步移将 N-乙酰胞壁酰-L-丙氨酸酰胺酶基因（cwlB；主要枯草芽孢杆菌自溶素）的上游区域克隆到大肠杆菌中。该区域的测序显示存在两个开放阅读框，一个（指定为cwbA）从UUG密码子开始，编码705个氨基酸的多肽，M（r）为76,725，另一个（指定为lppX）位于cwbA的上游，包含102个氨基酸并具有脂蛋白的信号序列特征。纯化CwbA蛋白并测定其N端氨基酸序列表明，其含有推测的信号肽，该信号肽是在N端第25位Ala之后加工的，并且成熟形式的M(r)为75,000。发现CwbA的N端和C端区域的氨基酸序列分别与CwlB的细胞壁结合结构域和spoIID基因产物的氨基酸序列高度同源。 CwbA 在体外将主要自溶素活性刺激大约三倍。这些数据表明 CwbA 是 Herbold, D. R. & Glaser, L. (1975; Journal of Biological Chemistry 250, 1676-1682) 报道的主要自溶素的修饰蛋白。 lppX 和 lacZ 基因之间的框内融合表明 lppX 在体内翻译并在指数生长期表达。
The upstream region of the N-acetylmuramoyl-L-alanine amidase gene (cwlB; a major Bacillus subtilis autolysin) was cloned into Escherichia coli by chromosome walking. Sequencing of the region showed the presence of two open reading frames, one (designated as cwbA) which starts at a UUG codon and encodes a polypeptide of 705 amino acids with an M(r) of 76,725, and the other (designated as lppX), upstream of cwbA, comprising 102 amino acids and having a signal sequence characteristic of a lipoprotein. Purification of the CwbA protein and determination of its N-terminal amino acid sequence revealed that it contains a presumed signal peptide which is processed after Ala at position 25 from the N-terminal, and that the M(r) of the mature form is 75,000. The amino acid sequences of the N-terminal and C-terminal regions of CwbA were found to be highly homologous with those of the cell wall binding domain of CwlB and the spoIID gene product, respectively. CwbA stimulated the major autolysin activity approximately threefold in vitro. These data indicate that CwbA is the modifier protein of the major autolysin reported by Herbold, D. R. & Glaser, L. (1975; Journal of Biological Chemistry 250, 1676-1682). In-frame fusion between the lppX and lacZ genes demonstrated that lppX is translated in vivo and expressed during the exponential growth phase.
影响因子：
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作者：
H. C. Wu;M. Tokunaga
通讯作者：
H. C. Wu;M. Tokunaga