葡萄球菌A蛋白基因的克隆及其IgG受体区新型表达载体的构建

Cloning of Protein A Gene from Staphylococcus aureus and Constructing a New Expression Vectors of the Domain of IgG-Fc Receptor

旨在克隆SPA基因并将该基因的IgG结合区亚克隆至毕赤酵母表达载体中。以金黄色葡萄球菌CowanI菌株基因组为模板,对葡萄球菌A蛋白(SPA)基因全长序列进行PCR扩增,再将PCR产物克隆入pMD18-T质粒,将DNA测序所得的结果用Blastn软件进行在线同源比对,经鉴定为SPA基因序列后,在线对其进行功能区域(IgG-Fc受体区)的预测,将该区域亚克隆入表达载体pPICZaA。结果显示,从CowanI菌株中成功地扩增到SPA基因,与NCBI中公布的序列同源性高达97%。

It was to construct the expression vector of the IgG receptor.The protein A gene of Staphylococcus aureus CowanI strain（SPA）was amplified by polymerase chain reaction in this study.The amplified fragment was cloned into the pMD18-T vector and sequenced,and the aim sequence of SPA was analyzed by blastn search,the function domains of which was predicted by the smart soft and suncloned into the expression vector of pPICZaA.Result showed that the amplified sequence is high homologous to the sequence of the protein A from the NCBI and the domain of IgG-Fc receptor from the amplified sequence was obtained.Therefore,SPA gene is amplified from the CowanI strain,and expression vector of the domain of IgG-Fc receptor was constructed successfully.