Herchenröder O, Turek R, Neumann-Haefelin D, Rethwilm A, Schneider J
Abteilung Virologie, Universität Freiburg, Germany.
Virology. 1995 Dec 20;214(2):685-9. doi: 10.1006/viro.1995.0086.
Infectious proviral clones of simian foamy virus isolated from chimpanzee (SFVcpz) were generated by long PCR. Two overlapping fragments representing the complete provirus were amplified from genomic DNA of infected cells. Four 8.8-kbp amplimers extending from base 1 of the provirus into the env gene and five 4.45-kbp amplimers reaching from env to the end of the 3'-LTR were cloned into pCR II. Subsequently, the proviral fragments were combined in a chessboard manner to generate 20 plasmids containing full-length proviral DNA. Four plasmids produced infectious virus after transfection of susceptible cells. A distinct proviral form bearing a deletion in the transactivator gene joining both exons of a second regulatory gene present in wild-type foamy virus-infected cells started to emerge 48 hr after transfection of BHK cells with infectious SFVcpz DNA. This observation supports a novel hypothesis to explain establishment of foamy virus latency. The transactivator protein Taf of SFVcpz transcomplemented for the homologous protein Bel-1 of the unique human foamy virus isolate (HFV) and Bel-1 exhibited the reciprocal activity, suggesting that HFV could represent a variant of chimpanzee foamy virus.
通过长链聚合酶链式反应(PCR)构建了从黑猩猩分离出的猿猴泡沫病毒(SFVcpz)的感染性前病毒克隆。从感染细胞的基因组DNA中扩增出代表完整前病毒的两个重叠片段。将四个从病毒前体的第1个碱基延伸至env基因的8.8千碱基对(kbp)扩增子和五个从env基因延伸至3'-长末端重复序列(LTR)末端的4.45 kbp扩增子克隆到pCR II中。随后,将前病毒片段以棋盘式组合,生成20个包含全长前病毒DNA的质粒。四个质粒在转染易感细胞后产生了感染性病毒。在用感染性SFVcpz DNA转染BHK细胞48小时后,一种在野生型泡沫病毒感染细胞中存在的连接第二个调节基因两个外显子的反式激活基因中带有缺失的独特前病毒形式开始出现。这一观察结果支持了一个新的假说来解释泡沫病毒潜伏期的建立。SFVcpz的反式激活蛋白Taf对独特的人类泡沫病毒分离株(HFV)的同源蛋白Bel-1具有反式互补作用,且Bel-1表现出相反的活性,这表明HFV可能是黑猩猩泡沫病毒的一个变体。