金黄色葡萄球菌TRAP基因的克隆与序列分析

参考Genbank已经发表的关于金黄色葡萄球菌TRAP蛋白的基因序列,设计合成1对引物,从本实验室保存的金黄色葡萄球菌Newman株中提取细菌DNA,对TRAP蛋白基因进行PCR扩增,产物经琼脂糖凝胶电泳分析,呈现1条约500bp的条带,回收纯化后,将其克隆至pMD18-T质粒载体中,进行核苷酸序列分析,然后与报道的TRAP蛋白基因进行比较。结果表明:所扩增的基因序列与报道的TRAP核苷酸的同源性高达100/,证实为金黄色葡萄球菌Newman株TRAP蛋白基因。

Cloning and Sequence Analysis of the TRAP Gene of Staphylococcus aureus

A pair of primer for TRAP (target of RNAIII Activating Protein) of staphylococcus aureus (S.aureus) was designed and synthesized according to the TRAP gene sequences of S aureus published in Genbank. After the extraction of DNA from the S.aureus Newman strain, the primers were used to amplify the TRAP gene, and the amplified product was analyzed through agar gel electrophoresis. It was shown that an about 500bp fragment was successfully amplified. Then, the 500bp fragment was linked to pMD18-T plasmid and was determined by sequence analysis. The result showed that there was an identity of 100/ between the amplified gene and the published TRAP gene and indicated that the amplified gene was one gene of Newman strain's TRAP.