中文摘要：

目的采用Shotgun法对牙龈卟啉单胞菌Pg301外膜蛋白进行分析,为筛选牙周炎疫苗候选抗原奠定基础。方法培养Pg301,提取外膜蛋白,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离蛋白,胶内酶解成多肽混合物,经高效液相色谱-串联质谱分析,结合美国国立生物技术信息中心蛋白质数据库进行蛋白鉴定,获得外膜蛋白成分。结果分析出586个唯一肽段,鉴定出93种蛋白,其中有50种是已确定的蛋白。结论 Shotgun法能够快速高效分析细菌外膜蛋白,为筛选保护性抗原、毒力相关候选蛋白等奠定基础。

英文摘要：

Objective Apply ＂Shotgun＂ method to analyze the outer membrane proteins （OMP） of Pg301 and set foundation for sieving candidate antigens of periodontitis vaccine. Methods Pg301 was cultured and OMP were extrac- ted, then separated by SDS-PAGE. Using trypsin to hydrolysis proteins in SDS-PAGE gel, so that peptide fragments could be acquired. Separate the peptides fragments using HPLC, and analyze every peptide fragment in MS/MS, so the sequence of each peptide could be obtained. With the help of SEQUEST software, searching the NCBI data bank, the proteins could be identified. Results Five hundred and eighty-six pcptides were analyzed and 93 proteins were ob- tained, in which 50 proteins had been confirmed. Conclusion More OMP element can be analyzed quickly and effec- tively by Shotgun method, which sets foundation for sieving protective antigens and candidate proteins that are related to virulence.