ABSTRACT
ObjectiveTo develop a transgenic mouse model for N.gonorrhoeae researches.Methods Human carcinoembryonic antigen-related cellular adhesion molecules 1 (hCEACAM1) eukaryotic expression vector,pCDPGICAM1,was used to generate transgenic mice by microinjection.The funder mice were screened by PCR,sequence analysis,Western blot and fluorescence-activated cell sorting analysis,respectively.The transgenic mice expressing hCEACAM1 were inoculated with N.gonorrhoeae intravaginally.Adhesion and infection of gonococci to mice were analyzed by bacteria culture and microscopy.Results Four (lines 50,53,54,and 59) of the 22 F0 generation transgenic mice were found to carry the transgene.The hCEACAM1 protein was expressed on the cell membrane of various tissues in the line 53 transgenic mouse.Compared with normal mice,N.gonorrhoeae can successfully infect and cause inflammation in the transgenic mice.Conclusion The hCEACAM1 transgenic mouse can be used as an animal model for gonococcal infections.
ABSTRACT
Objective To study the role of human carcinoembryonic antigen-related cellular adhesion molecule 1 (hCEACAM1)in mediating the specific adhesion of N. gonorrhoeae to its human host cells.Methods A recombinant eukaryotic expression vector pCDPGICEA1 was constructed by putting hCEACAM1 cDNA behind both hCD46 promoter and rabbit β-globulin intron 2,and with which,the COS-1 cells were transfected. Following G418 selection, the COS-1 cells expressing hCEACAM1 were sorted out with flow cytometry. The adhesion of N. gonorrhoeae to the gene transfected COS-1 cells was analyzed with bacterial binding assay. Results hCEACAM1 cDNA could be expressed effectively under the direction of hCD46 promoter and rabbit β-globulin intron 2,and N. gonorrhoeae could adhere to COS-1 cells expressing hCEACAM1. Conclusion hCEACAM1 can mediate the adhesion of N. gonorrhoeae to animal originated COS-1 cells. thus its transgenic mice may be used as a novel animal model for studying N. gonorrhoeae infection.