Perricaudet M, Le Moullec J M, Pettersson U
Proc Natl Acad Sci U S A. 1980 Jul;77(7):3778-82. doi: 10.1073/pnas.77.7.3778.
Early adenovirus type 2(Ad2) mRNA sequences have been cloned by using the pBR322 plasmid as a vector. Two clones that include sequences from region E1B were identified and their DNAs were characterized by hybridization, restriction enzyme cleavage, and DNA sequence analysis. The results showed that the clones were derived from two different spliced mRNAs. By combining our results with the established DNA sequence for region E1B of the closely related adenovirus type 5[Maat, J., van Beveren, C.P. & van Ormondt, H. (1980) Gene, in press] it was possible to deduce the structure of a 13S and a 22S mRNA. The two mRNAs differ from each other by the size of their intervening sequences. If translation starts at the first AUG following the cap, the 22S mRNA encodes a Mr 67,000 polypeptide that is terminated by a UGA stop codon located immediately before the splice, whereas the 13S mRNA encodes a Mr 20,000 polypeptide that is translated in different reading frames before and after the splice. The Mr 20,000 and 67,000 polypeptides correspond in molecular weight to two proteins that invariably are precipitated from infected cell extracts by antisera from animals carrying adenovirus-induced tumors.
利用pBR322质粒作为载体克隆了早期2型腺病毒(Ad2)的mRNA序列。鉴定出两个包含E1B区序列的克隆,并通过杂交、限制性内切酶切割和DNA序列分析对其DNA进行了表征。结果表明,这些克隆来自两种不同的剪接mRNA。通过将我们的结果与密切相关的5型腺病毒E1B区已确定的DNA序列相结合[马特,J.,范贝弗伦,C.P.和范奥蒙德特,H.(1980年)《基因》,即将发表],有可能推断出13S和22S mRNA的结构。这两种mRNA的不同之处在于它们间隔序列的大小。如果翻译从帽子之后的第一个AUG开始,22S mRNA编码一种分子量为67,000的多肽,该多肽由紧接在剪接之前的UGA终止密码子终止,而13S mRNA编码一种分子量为20,000的多肽,该多肽在剪接前后以不同的阅读框进行翻译。分子量为20,000和67,000的多肽在分子量上与两种蛋白质相对应,这两种蛋白质总是被携带腺病毒诱导肿瘤的动物的抗血清从感染细胞提取物中沉淀出来。