淋病奈瑟菌对hCD46基因转染COS-1细胞的黏附

为研究hCD46在菌毛介导的淋病奈瑟菌宿主特异性黏附过程中的作用。用连接PCR技术扩增出启动子与cDNA相连的hCD46小基因，并将其置换出pcDNA3.1载体中的CMV启动子，构建成重组真核表达载体pCD46。转染COS-1细胞后用间接免疫荧光法检测到hCD46 cDNA在其自身启动子的指导下可在COS-1细胞膜表面有效表达，Western blotting检测表明表达产物的分子正确，用流式细胞术分选表达hCD46的基因转染细胞COS-1-46。细菌黏附实验显示菌毛阳性淋病奈瑟菌临床分离株对COS-1-46细胞有较强的黏附性。提示hCD46是菌毛介导的淋病奈瑟菌特异性黏附于人黏膜细胞的一种重要受体，hCD46小基因可用于淋病奈瑟菌感染转基因小鼠模型的制备。

Adherence of Neisseria gonorrhoeae to hCD46 expressing COS-1 cells

To study the role of hCD46 in mediating the specific adhesion by N. gonorrhoeae to its human host, an hCD46 minigene including the promoter and cDNA was synthesized by using ligation-mediated PCR, and cloned into vector pcDNA3.1 replacing the original CMV promoter. The resultant recombinant vector pCD46 was transfected into COS-1 cells. The effective expression of hCD46 on the membrane of COS-1 cells was detected by indrect immunofluorscence assay; Western blot revealed that the molecular weight of the recombinant hCD46 was correct. The hCD46 expressing COS-1-46 cells were sorted by FACS and could be adhered by piliated N. gonorrhoeae stain YZ1228 revealed by bacteria binding assay. These data indicate that hCD46 is a kind of crucial pilus receptor that mediates the host specific adhesion by N. gonorrhoeae, and the hCD46 minigene, pCD46, can be used to develop transgenic mice model for the study of gonococcal diseases.