Cloning and purification of a unique lysozyme produced by Bacillus phage phi 29.
Cloning and purification of a unique lysozyme produced by Bacillus phage phi 29.
复制标题
芽孢杆菌噬菌体 phi 29 产生的独特溶菌酶的克隆和纯化。
DOI：
10.1073/pnas.84.4.955
复制
发表时间：
1987
影响因子：
11.1
通讯作者：
Ito,J
中科院分区：
文献类型：
--
作者：
Saedi,MS;Garvey,KJ;Ito,J
噬菌体 phi 29 染色体的 DNA 片段编码 phi 29 基因 15 的整个序列，已被克隆到大肠杆菌表达载体 pPLc245 中，处于噬菌体 lambda 主要向左启动子 PL 的控制下。热诱导后，表观分子量为 26 kDa 的蛋白质被过量产生。该蛋白质的分子量对应于根据其核苷酸序列预测的基因 15 产物的 28 kDa。过量产生的蛋白质已被纯化至接近均质，并通过其N末端的氨基酸序列分析证实是基因15的产物。基因15的纯化产物具有与其他噬菌体型溶菌酶类似的溶菌酶活性：噬菌体T4基因e和噬菌体P22基因19的产物。然而，据我们所知，phi 29溶菌酶在结构上在噬菌体型溶菌酶中是独特的。
A DNA fragment of the bacteriophage phi 29 chromosome, encoding the entire sequence of phi 29 gene 15, has been cloned into the Escherichia coli expression vector pPLc245 under the control of the phage lambda major leftward promoter, PL. Upon heat induction, a protein with an apparent molecular mass of 26 kDa was overproduced. The molecular mass of this protein corresponds to the 28 kDa predicted for the product of gene 15 from its nucleotide sequence. The overproduced protein has been purified to near homogeneity and confirmed to be the product of gene 15 by amino acid sequence analysis of its N terminus. The purified product of gene 15 has a lysozyme activity similar to other phage-type lysozymes: products of phage T4 gene e and of phage P22 gene 19. However, to our knowledge phi 29 lysozyme is structurally unique among the phage-type lysozymes.