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PRCII肉瘤病毒基因组和鸡原癌基因c-fps的分子克隆

Molecular cloning of the PRCII sarcoma viral genome and the chicken proto-oncogene c-fps.

作者信息

Hammond C I, Vogt P K, Bishop J M

出版信息

Virology. 1985 May;143(1):300-8. doi: 10.1016/0042-6822(85)90117-5.

Abstract

The class II avian sarcoma viruses comprise PRCII, PRCIIp, PRCIV, URI, 16L, and Fujinami. The members of this class are all replication-defective viruses containing various amounts of a transforming sequence called v-fps. PRCII contains the smallest amount of fps-specific sequences, transforms fibroblasts in tissue culture, but is only weakly tumorigenic. As a first step in understanding variations in pathogenicity among the class II avian sarcoma viruses and the mechanism by which the oncogene of these viruses was transduced from a single cellular locus, we have molecularly cloned the viral genome of PRCII, its related helper PRCII-AV, and the chicken proto-oncogene (c-fps) from which v-fps derived. The fps-specific region within the cloned PRCII genome was shown to be 0.8-1.0 kb smaller than that of the Fujinami fps-specific region, in agreement with previous studies. Transfection of the cloned DNAs into primary chicken cells demonstrated that both clones (PRCII and PRCII-AV) are biologically active. The cloned PRCII genome is helper dependent and produces a gag-fusion phosphoprotein (P105) which is phosphorylated on a tyrosine residue. The cloned PRCII-AV genome produces infectious virus and can function as a helper for the cloned PRCII genome in transfection assays. Three overlapping recombinant lambda clones homologous to v-fps from a chicken genomic library have been isolated. One of these, lambda-c-fps(2), contains all of the cellular sequences homologous to v-fps. In the aggregate, the three molecular clones may represent the entirety of c-fps.

摘要

II类禽肉瘤病毒包括PRCII、PRCIIp、PRCIV、URI、16L和藤浪(Fujinami)病毒。该类病毒成员均为复制缺陷型病毒,含有不同量的一种名为v-fps的转化序列。PRCII所含的fps特异性序列量最少,能在组织培养中转化成纤维细胞,但致瘤性较弱。作为了解II类禽肉瘤病毒致病性差异以及这些病毒的癌基因从单个细胞位点转导机制的第一步,我们已对PRCII病毒基因组、其相关辅助病毒PRCII-AV以及v-fps所源自的鸡原癌基因(c-fps)进行了分子克隆。克隆的PRCII基因组中的fps特异性区域比藤浪病毒fps特异性区域小0.8 - 1.0 kb,这与先前的研究一致。将克隆的DNA转染到原代鸡细胞中表明,两个克隆(PRCII和PRCII-AV)均具有生物学活性。克隆的PRCII基因组依赖辅助病毒,产生一种在酪氨酸残基上磷酸化的gag融合磷蛋白(P105)。克隆的PRCII-AV基因组产生传染性病毒,并且在转染试验中可作为克隆的PRCII基因组的辅助病毒。已从鸡基因组文库中分离出三个与v-fps同源的重叠重组λ克隆。其中一个,λ-c-fps(2),包含所有与v-fps同源的细胞序列。总体而言,这三个分子克隆可能代表了c-fps的全部。

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