Jayaswal R K, Lee Y I, Wilkinson B J
Department of Biological Sciences, Illinois State University, Normal 61761.
J Bacteriol. 1990 Oct;172(10):5783-8. doi: 10.1128/jb.172.10.5783-5788.1990.
A gene of Staphylococcus aureus PS47 encoding lytic activity was cloned and expressed in Escherichia coli. Deletion analysis of a recombinant plasmid carrying a 7.4-kilobase-pair fragment (kbp) of S. aureus DNA suggested that the gene was located within a 2.5-kbp EcoRI-XbaI fragment. Analysis of extracts of E. coli harboring recombinant plasmids on denaturing polyacrylamide gels containing purified cell walls of S. aureus showed a clearing zone by a polypeptide of apparent Mr 23,000. The release of dinitrophenylalanine but not reducing groups from purified cell walls by a cell extract of recombinant E. coli suggested that we had cloned an N-acetylmuramyl-L-alanine amidase.
克隆了金黄色葡萄球菌PS47编码溶菌活性的基因,并在大肠杆菌中表达。对携带金黄色葡萄球菌DNA 7.4千碱基对片段(kbp)的重组质粒进行缺失分析表明,该基因位于一个2.5 kbp的EcoRI - XbaI片段内。在含有纯化的金黄色葡萄球菌细胞壁的变性聚丙烯酰胺凝胶上分析携带重组质粒的大肠杆菌提取物,结果显示一条表观分子量为23,000的多肽形成了一个透明区。重组大肠杆菌的细胞提取物能从纯化的细胞壁中释放二硝基苯丙氨酸,但不能释放还原基团,这表明我们克隆的是一种N - 乙酰胞壁酰 - L - 丙氨酸酰胺酶。