Rethwilm A, Baunach G, Netzer K O, Maurer B, Borisch B, ter Meulen V
Institut für Virologie, Universität Würzburg, FRG.
Nucleic Acids Res. 1990 Feb 25;18(4):733-8. doi: 10.1093/nar/18.4.733.
An infectious molecular clone (pHSRV) of the human Spumaretrovirus (HSRV) was constructed using viral DNA and cDNA clones. The infectivity of pHSRV was proven by transfection of cell cultures and subsequent infection of susceptible cultures with cell free transfection derived virus. pHSRV derived virus produced foamy virus typical cytopathic effects in susceptible cultures. Infected cells could be stained specifically with foamy virus antisera by means of indirect immunofluorescence. Radioimmunoprecipitation revealed the presence of characteristic HSRV structural proteins in pHSRV infected cultures. By cotransfection of pHSRV and an indicator plasmid it was found that pHSRV is able to transactivate the viral LTR. Viral transcripts were found to be approximately 200 bases longer in pHSRV infected cultures compared to wildtype infected cultures. This difference is most likely due to an insertion of DNA of non-viral origin in the U3 region of the 3'LTR of the infectious clone.
利用病毒DNA和cDNA克隆构建了人类泡沫逆转录病毒(HSRV)的感染性分子克隆(pHSRV)。通过转染细胞培养物以及随后用无细胞转染衍生病毒感染易感培养物,证明了pHSRV的感染性。pHSRV衍生病毒在易感培养物中产生了泡沫病毒典型的细胞病变效应。通过间接免疫荧光法,感染细胞可用泡沫病毒抗血清进行特异性染色。放射免疫沉淀显示在pHSRV感染的培养物中存在特征性的HSRV结构蛋白。通过共转染pHSRV和一个指示质粒,发现pHSRV能够反式激活病毒长末端重复序列(LTR)。与野生型感染的培养物相比,在pHSRV感染的培养物中发现病毒转录本大约长200个碱基。这种差异很可能是由于在感染性克隆的3'LTR的U3区域插入了非病毒来源的DNA。