伤寒沙门氏菌外膜蛋白的分离和鉴定

用 Triton X-100处理伤寒沙门氏菌外膜,经3次超离后从上清液中获得外膜蛋白(OMPs)。SDS-PAGE 显示,OMPs 蛋白带的分子量在17～70KD 间,其中分子量为36～41KD的主要蛋白占总 OMPs 的50～60%。经测定2-酮-3-脱氧辛酸(KDO)的含量间接测得,OMPs 中脂多糖(LPS)的污染量约为4%。用 OMPs 免疫家兔,抗体效价琼脂双扩散法为1:64,ELISA 法为1:12 500。免疫印迹法显示,兔抗-OMP 血清主要与 OMPs 中的主要蛋白发生反应。

Isolation and Identification for OMPs of Salmonella Typhi

Abstract Following treating outer membrane of the Salmonella typhi with nonionic detergent Triton X-100,outer membrane proteins(OMPs)were obtained from the super- natant fraction after three times ultracentrifugation.SDS-PAGE patterns of OMPssh- owed protein bands with molecular weight ranging from 17 to 70 KD.The major grou- ps of proteins with molecular weight between 36～41 KD occupy 50～60% of total OM- Ps.OMPs were found to be contaminated with approximately 4% lipopolysaccharide as determined indirectly by measuring the concentration of 2-keto-3-deoxyoctulosonic acid(KDO).Rabbit antiserum to OMPs were assayed by double immunodiffusion and ELISA.The antibody titers were 1:64 and 1:12500 respectively.Western blotting analys- is showed that rabbit antiserum to OMPs reacted mainly with the major Proteins in OMPs.