CAJ | 학술논문

目的：制备携带增强绿色荧光蛋白（ EGFP）基因和ENV包膜蛋白的HIV-1 B亚型假病毒用于感染研究，并建立可行的鉴定方法。方法双质粒共转染HEK293 T细胞后收获病毒上清， TRIzol法提取病毒基因组并采用RT-PCR进行报告基因扩增，Western印迹和ELISA法检测病毒P24抗原。假病毒感染HIV-1允许细胞，进行报告基因检测、病毒滴度测定以及单轮感染活性实验研究。结果与结论建立了HIV-1假病毒制备与鉴定方法，制备获得了B型HIV-1假病毒，经鉴定具备感染SupT1和TZM-bl细胞能力，为HIV-1与宿主细胞相互作用研究奠定了基础。
목적：제비휴대증강록색형광단백（ EGFP）기인화ENV포막단백적HIV-1 B아형가병독용우감염연구，병건립가행적감정방법。방법쌍질립공전염HEK293 T세포후수획병독상청， TRIzol법제취병독기인조병채용RT-PCR진행보고기인확증，Western인적화ELISA법검측병독P24항원。가병독감염HIV-1윤허세포，진행보고기인검측、병독적도측정이급단륜감염활성실험연구。결과여결론건립료HIV-1가병독제비여감정방법，제비획득료B형HIV-1가병독，경감정구비감염SupT1화TZM-bl세포능력，위HIV-1여숙주세포상호작용연구전정료기출。
Objective To prepare the Env-pseudotyped subtype B HIV-1 with enhanced green fluorescent protein ( EG-FP) gene,explore HIV-1 infection mechanisms and develop feasible methods of identification .Methods The Env-pseudo-typed viruses were packaged in HEK293T cells by cotransfection, and the reporter gene and P24 protein were detected by PCR, Western blot and ELISA .Reporter gene amplification , viral titration assay and a single round of infection assay were performed after the env-pseudotyped viruses infected HIV-1 permissive cell .Results and Conclusion A generation and identification method of the pseudotyped HIV-1 was established . The Env-pseudotyped subtype B HIV-1 has been prepared, which is able to infect SupT1 and TZM-bl cells through infection assay .