Srivatsan E S, Deininger P L, Friedmann T
J Virol. 1981 Jan;37(1):244-7. doi: 10.1128/JVI.37.1.244-247.1981.
Double-stranded DNA complementary to total cytoplasmic polyadenylated RNA isolated late in infection from polyoma virus-infected mouse 3T6 cells was cloned in Escherichia coli by using the large HindIII-BamHI fragment of pBR322 plasmid DNA. Polyoma-specific DNA inserts were detected by hybridization, and then nucleotide sequences were determined from two clones. The sequence of the (formula see text) with prototypical mammalian splice sites, and the dashed arrows indicate possible alternative splice sites leading to the same spliced product. A sequence of 897 nucleotides was spliced out of the primary transcript during the processing of the mature VP1 mRNA. Restriction enzyme mapping with four other independently isolated clones indicates that these are the major splicing signals for the VP1 message. The distal splice site is 48 nucleotides upstream from the initiator codon.
与从多瘤病毒感染的小鼠3T6细胞感染后期分离的总细胞质聚腺苷酸化RNA互补的双链DNA,通过使用pBR322质粒DNA的大HindIII - BamHI片段克隆到大肠杆菌中。通过杂交检测多瘤病毒特异性DNA插入片段,然后从两个克隆中确定核苷酸序列。(公式见文本)具有典型哺乳动物剪接位点的序列,虚线箭头表示导致相同剪接产物的可能替代剪接位点。在成熟VP1 mRNA的加工过程中,897个核苷酸的序列从初级转录本中剪接出来。用其他四个独立分离的克隆进行限制性酶切图谱分析表明,这些是VP1信使的主要剪接信号。远端剪接位点在起始密码子上游48个核苷酸处。