Shibuya M, Wang L H, Hanafusa H
J Virol. 1982 Jun;42(3):1007-16. doi: 10.1128/JVI.42.3.1007-1016.1982.
Full-length proviral DNA of Fujinami sarcoma virus (FSV) of chickens was molecularly cloned and characterized. An analysis of FSV DNA integrated in mammalian cells showed that restriction endonuclease SacI has a single cleavage site on FSV DNA. Unintegrated closed circular FSV DNA obtained from newly infected cells was linearized by digestion with SacI and cloned into lambdagtWES.lambdaB. The following three different molecules were isolated: FSV-1 (4.4 kilobases [kb]) and FSV-2 (4.7 kb), which appeared to be full-length FSV DNA molecules containing either one or two copies of the long terminal repeat structure, and FSV-3 (6 kb), which consisted of part FSV DNA and part DNA of unknown origin. An analysis of the structure of cloned FSV-1 and FSV-2 DNA molecules by restriction endonuclease mapping and hybridization with appropriate probes showed that about 2.6 kb of the FSV-unique sequence called FSV-fps is located in the middle of the FSV genome and is flanked by helper virus-derived sequences of about 1.3 kb at the 5' end and 0.5 kb at the 3' end. The long terminal repeats of FSV were found to have no cleavage site for either EcoRI or PvuI. Upon transfection, both FSV-1 DNA and FSV-2 DNA were able to transform mammalian fibroblasts. Four (32)P-labeled DNA fragments derived from different portions of the FSV-fps sequence were used for hybridization to viral RNAs. We found that sequences within the 3' half of the FSV-fps gene are homologous to RNAs of PRCII avian sarcoma virus and the Snyder-Theilen strain of feline sarcoma virus, both of which were previously shown to contain transforming genes related to FSV-fps. These results suggest that the 3' portion of the FSV-fps sequence may be crucial for the transforming activity of fps-related oncogenic sequences.
鸡的 Fujinami 肉瘤病毒（FSV）的全长前病毒 DNA 进行了分子克隆和特性分析。对整合在哺乳动物细胞中的 FSV DNA 的分析表明，限制性内切酶 SacI 在 FSV DNA 上有一个单一的切割位点。从新感染细胞中获得的未整合的闭环 FSV DNA 经 SacI 消化后线性化，并克隆到 lambdagtWES.lambdaB 中。分离出以下三种不同的分子：FSV-1（4.4 千碱基 [kb]）和 FSV-2（4.7 kb），它们似乎是包含一个或两个长末端重复结构拷贝的全长 FSV DNA 分子，以及 FSV-3（6 kb），它由部分 FSV DNA 和部分未知来源的 DNA 组成。通过限制性内切酶图谱分析和与适当探针杂交对克隆的 FSV-1 和 FSV-2 DNA 分子结构进行分析表明，约 2.6 kb 的称为 FSV-fps 的 FSV 独特序列位于 FSV 基因组的中部，在 5' 端两侧是约 1.3 kb 的辅助病毒衍生序列，在 3' 端两侧是约 0.5 kb 的辅助病毒衍生序列。发现 FSV 的长末端重复序列对 EcoRI 或 PvuI 均无切割位点。转染后，FSV-1 DNA 和 FSV-2 DNA 均能够转化哺乳动物成纤维细胞。来自 FSV-fps 序列不同部分的四个（32）P 标记的 DNA 片段用于与病毒 RNA 杂交。我们发现 FSV-fps 基因 3' 半部分内的序列与 PRCII 禽肉瘤病毒和 Snyder-Theilen 株猫肉瘤病毒的 RNA 同源，这两种病毒先前已被证明含有与 FSV-fps 相关的转化基因。这些结果表明，FSV-fps 序列的 3' 部分可能对 fps 相关致癌序列的转化活性至关重要。
