Fasel N, Pearson K, Buetti E, Diggelmann H
EMBO J. 1982;1(1):3-7. doi: 10.1002/j.1460-2075.1982.tb01115.x.
Starting from a biologically active recombinant DNA clone of exogenous unintegrated GR mouse mammary tumor virus, we have generated three subclones of PstI fragments of 1.45, 1.1, and 2.0 kb in the plasmid vector PBR322. The nucleotide sequence has been determined for the clone of 1.45 kb which includes almost the complete region of the long terminal repeat (LTR) plus an adjacent stretch of unique sequence DNA. A short region of the 2.0 kb clone, containing the beginning of the LTR, has also been sequenced. Starting with the A of an initiation codon outside the LTR, we detected an open reading frame of 960 nucleotides, potentially coding for a protein of 320 amino acids (36K). Two hundred nucleotides downstream from the termination codon, and approximately 25 nucleotides upstream from the presumptive initiation site of viral RNA synthesis, we found a promoter-like sequence. The sequence AGTAAA was detected approximately 15-20 nucleotides upstream from the 3' end of virion RNA and probably serves as a polyadenylation signal. The 1.45 kb PstI fragment has been transfected into Ltk- cells together with a plasmid containing the thymidine kinase gene of herpes simplex virus. The virus-specific RNA synthesis detected in a Tk+ cell clone was strongly stimulated by the addition of dexamethasone.
从外源性未整合的GR小鼠乳腺肿瘤病毒的生物活性重组DNA克隆开始,我们在质粒载体PBR322中产生了1.45、1.1和2.0 kb的PstI片段的三个亚克隆。已确定了1.45 kb克隆的核苷酸序列,该克隆包括几乎完整的长末端重复序列(LTR)区域以及一段相邻的独特序列DNA。还对2.0 kb克隆的一个短区域进行了测序,该区域包含LTR的起始部分。从LTR外起始密码子的A开始,我们检测到一个960个核苷酸的开放阅读框,可能编码一个320个氨基酸(36K)的蛋白质。在终止密码子下游200个核苷酸处,以及在病毒RNA合成假定起始位点上游约25个核苷酸处,我们发现了一个类似启动子的序列。在病毒粒子RNA的3'端上游约15 - 20个核苷酸处检测到序列AGTAAA,它可能作为聚腺苷酸化信号。1.45 kb的PstI片段已与含有单纯疱疹病毒胸苷激酶基因的质粒一起转染到Ltk-细胞中。在一个Tk+细胞克隆中检测到的病毒特异性RNA合成受到地塞米松添加的强烈刺激。