金黄色葡萄球菌基因组表达文库的构建及免疫学初步筛选

Construction and Preliminary Immunoscreening of Genome Expression Library of Staphylococcus aureus

为从基因组水平筛选金黄色葡萄球菌疫苗抗原，建立金黄色葡萄球菌基因组表达文库，用患者恢复期血清对表达文库进行血清学筛选，并对获得的阳性克隆进行序列测定及同源性分析。结果显示，构建的金黄色葡萄球菌基因组表达文库共有5x10。个克隆，片段插入正确率为91．67％。对文库进行初步筛选，共获得6个阳性克隆，经测序鉴定和同源性分析发现6个克隆涉及金黄色葡萄球菌基因组6个开放阅读框（ORF）。结果表明，本研究建立的疫苗抗原筛选方法可用于金黄色葡萄球菌疫苗抗原的系统筛选。

In order to achieve genome-wide screening of vaccine antigens of Staphylococcus aureus, genome expression library of Staphylococcus aureus was constructed and the expression library was immunoscreened with human convalescent sera. The obtained positive clones were sequenced and analyzed. The result showed that reservoir capacity of the constructed expression library was 5 x 104 clones and the insertion efficiency was 91.67%. Six positive clones were obtained after preliminary immunoscreening. After sequencing and bioinformatics analysis, six open reading frames （ORFs） were confirmed in the six clones. In conclusion, the established immunoscreening of vaccine antigens in this study based on genome expression library has the potential for systematic screening of vaccine antigens from Staphylococcus aureus.