Mitsialis S A, Caplan S, Guntaka R V
Mol Cell Biol. 1983 Nov;3(11):1975-84. doi: 10.1128/mcb.3.11.1975-1984.1983.
The regulatory elements present in the long terminal repeat (LTR) of avian sarcoma virus DNA were analyzed by recombinant DNA techniques coupled with DNA-mediated gene transfer in avian as well as mammalian cells. For this purpose, the neomycin resistance gene from transposon Tn5 was inserted downstream from the avian sarcoma virus LTR, and the recombinant plasmid DNA was introduced into cells by the calcium phosphate technique. Cells resistant to the drug G-418 were selected. Analysis of the RNA transcripts made in vivo in these transformants indicated that initiation and termination of the transcripts occurred in the LTR sequences. Deletions were then introduced into the LTR, and their effect on transcription was also studied. These results allowed us to identify a strong regulatory sequence between nucleotides -299 and -114 in the LTR of avian sarcoma virus.
通过重组DNA技术结合DNA介导的基因转移,在禽类和哺乳动物细胞中分析了禽肉瘤病毒DNA长末端重复序列(LTR)中的调控元件。为此,将来自转座子Tn5的新霉素抗性基因插入禽肉瘤病毒LTR的下游,并通过磷酸钙技术将重组质粒DNA导入细胞。选择对药物G-418耐药的细胞。对这些转化体中体内产生的RNA转录本的分析表明,转录本的起始和终止发生在LTR序列中。然后在LTR中引入缺失,并研究其对转录的影响。这些结果使我们能够在禽肉瘤病毒LTR中鉴定出一个位于核苷酸-299和-114之间的强调控序列。