FBJ MURINE OSTEO-SARCOMA VIRUS - IDENTIFICATION AND MOLECULAR-CLONING OF BIOLOGICALLY-ACTIVE PROVIRAL DNA
FBJ MURINE OSTEO-SARCOMA VIRUS - IDENTIFICATION AND MOLECULAR-CLONING OF BIOLOGICALLY-ACTIVE PROVIRAL DNA
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DOI：
10.1128/jvi.44.2.674-682.1982
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发表时间：
1982-01-01
影响因子：
5.4
通讯作者：
VERMA, IM
中科院分区：
文献类型：
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作者：
CURRAN, T;PETERS, G;VERMA, IM
在 FBJ-MSV 转化的非生产大鼠细胞中鉴定出含有 FBJ 鼠骨肉瘤病毒 (FBJ-MSV) 前病毒 DNA 的 12.0-kilobase (kb) EcoRi 限制性片段，并在噬菌体 Charon 30 (λFBJ-1) 中进行分子克隆。从λFBJ-1中分离出含有完整FBJ-MSV原病毒DNA的5.8-kb HindIII片段，并随后亚克隆到质粒pBR322(pFBJ-2)中。重组质粒 pFBJ-2 的 DNA 能够诱导组织培养中大鼠成纤维细胞的形态转化。转染的细胞含有对 FBJ-MSV 转化的细胞特异的 p55 和 p39 抗原。通过限制性核酸内切酶图谱分析了 FBJ-MSV 原病毒的组织，并描绘了与辅助病毒非同源的区域。该区域特异的序列（可能是病毒 fos 基因）被亚克隆并用作探针，以鉴定来自多种哺乳动物物种的细胞的正常基因组中存在的相关序列（细胞 fos）。在 FBJ-MSV 转化细胞中鉴定出与病毒 fos 探针杂交的单一大小（3.4 kb 长）RNA 类。
A 12.0-kilobase (kb) 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. 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 kb long)class of RNA hybridizing to the viral fos probe was identified in FBJ-MSV-transformed cells.