【摘要】 根据GenBank上发表的金黄色葡萄球菌β-溶血素基因序列,设计了一对特异性引物。提取临床乳腺炎致病性金黄色葡萄球菌山东株zfb的全基因组DNA为PCR模板,扩增β-溶血素基因,将PCR产物纯化后连接到pMD18-T载体中。测序结果表明,扩增片断含有993 bp的ORF,可编码含330个氨基酸和1个终止子的成熟蛋白,与已报道的金黄色葡萄球菌β-溶血素蛋白的氨基酸组成同源性为99.4%。软件分析显示,克隆的金葡菌β-溶血素是一种依赖于金属离子的磷脂酶。上述结果显示,已获得金黄色葡萄球菌β-溶血素基因,为获得重组金黄色葡萄球菌β-溶血素蛋白及对其结构和功能的研究奠定了基础。更多还原

【Abstract】 One pair of specific primers of Staphylococcus aureus β-hemolysin gene was designed and synthesized according to the published sequence in GenBank.Staphylococcus aureus zfb was isolated and identified from the cows with mastitis in Shandong Province.β-hemolysin gene was cloned from genome DNA of Staphylococcus aureus zfb by PCR,and the PCR product was inserted into vector pMD18-T.Sequence analysis showed that the Staphylococcus aureus β-hemolysin gene was composed of 993 nucleotides encoding a mature polypeptide with 330 amino acids and one terminator.The comparison with other Staphylococcus aureus β-hemolysin gene showed that the highest amino acid identity was 99.4%.It showed that Staphylococcus aureus β-hemolysin was a Metal-dependent phospholipase by the software analysis.The results showed that the cloned plasmid was successfully constructed.更多还原