中国生物工程杂志

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.