Curran T, Peters G, Van Beveren C, Teich N M, Verma I M
J Virol. 1982 Nov;44(2):674-82. doi: 10.1128/JVI.44.2.674-682.1982.
A 12.0-kilobase EcoRI restriction fragment containing FBJ murine osteosarcoma virus (FBJ-MSV) proviral DNA was identified in FBJ-MSV-transformed nonproducer rat cells and molecularly cloned in bacteriophage Charon 30 (lambda FBJ-1). A 5.8-kb HindIII fragment containing the entire FBJ-MSV proviral DNA was isolated from lambda FBJ-1 and subsequently subcloned in plasmid pBR322 (pFBJ-2). The DNA from recombinant plasmid pFBJ-2 was able to induce morphological transformation of rat fibroblasts in tissue culture. Transfected cells contained the p55 and p39 antigens specific for cells transformed by FBJ-MSV (T. Curran and N. M. Teich, J. Virol. 42:114-122, 1982). The organization of the FBJ-MSV provirus was analyzed by restriction endonuclease mapping, and a region of nonhomology with the helper virus was delineated. Sequences specific for this region (presumably the viral fos gene) were subcloned and used as a probe to identify related sequences present in the normal genomes of cells from a variety of mammalian species (cellular fos). A single-size (3.4 kilobases long) class of RNA hybridizing to the viral fos probe was identified in FBJ-MSV-transformed cells.
在经FBJ小鼠骨肉瘤病毒(FBJ-MSV)转化的非生产性大鼠细胞中鉴定出一个含有FBJ-MSV前病毒DNA的12.0千碱基EcoRI限制性片段,并将其分子克隆到噬菌体Charon 30(λFBJ-1)中。从λFBJ-1中分离出一个含有完整FBJ-MSV前病毒DNA的5.8千碱基HindIII片段,随后将其亚克隆到质粒pBR322(pFBJ-2)中。重组质粒pFBJ-2的DNA能够在组织培养中诱导大鼠成纤维细胞的形态转化。转染细胞含有FBJ-MSV转化细胞特有的p55和p39抗原(T. Curran和N. M. Teich,《病毒学杂志》42:114 - 122,1982)。通过限制性内切酶图谱分析了FBJ-MSV前病毒的组织,并划定了与辅助病毒的非同源区域。将该区域特有的序列(推测为病毒fos基因)亚克隆,并用作探针来鉴定多种哺乳动物细胞正常基因组中存在的相关序列(细胞fos)。在FBJ-MSV转化细胞中鉴定出一类与病毒fos探针杂交的单一大小(3.4千碱基长)的RNA。