嵌合人/牛免疫缺陷病毒cDNA(pHBIV-2)传染性克隆的构建及其生物学活性  

Construction and Bioactivity Analysis of a pHBIV-2 Chimeric cDNA Clone

作　　者：杨怡姝[1] 陈国敏[2] 陈启民[3] 耿运琪[3] 曾毅[2] 

机构地区：[1]北京工业大学生命科学与生物工程学院,北京100022 [2]中国疾病预防控制中心病毒病预防控制所,北京100052 [3]南开大学生命科学学院,天津300071

出　　处：《病毒学报》2004年第2期138-142,共5页Chinese Journal of Virology

基　　金：国家重点基础研究发展规划项目(G1999054107)

摘　　要：为探讨牛免疫缺陷病毒(BIV)Tat能否在功能上取代HIVTat,构建用BIVtat取代HIVtat的嵌合人/牛免疫缺陷病毒(pHBIV 2)cDNA,将其转染人源MT4细胞。PCR、RT PCR法检测到嵌合基因组在MT4细胞中可稳定地存在并转录;套式Alu PCR法检测到嵌合基因组可整合到细胞基因组中;RTase活性测定及IFA检测显示,嵌合基因在MT4细胞中得到了翻译。结果表明,HIV的tat基因用BIVtat取代后产生的传染性cDNA克隆,仍能在人源MT4细胞中产生有复制性的重组病毒。In order to study whether BIV Tat could substitute HIV Tat,the chimeric human/bovine immunodeficiency virus(pHBIV-2)cDNA was constructed by replacing HIV tat with BIV tat, then transfected into human MT_4 cell.The stable existence and transcription of the chimerae in MT_4 cell were demonstrated by PCR and RT-PCR.The integration of chimerae into MT_4 cell was verified by nested Alu-PCR.Reverse transcriptase assay and IFA showed the chimeric genome had been expressed.In conclusion,BIV tat could substitute HIV tat in the chimerae and produce replicative recombinant HBIV-2 virus in MT_4 cell.

关 键 词：人免疫缺陷病毒 牛免疫缺陷病毒 反式激活因子 MT4细胞 CDNA BIV 

分 类 号：R373[医药卫生—病原生物学] Q78[医药卫生—基础医学]