Ahn B Y, Moss B
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
J Virol. 1989 Jan;63(1):226-32. doi: 10.1128/JVI.63.1.226-232.1989.
Evidence for capped poly(A) leaders of variable lengths located immediately upstream of the translation initiation codon was obtained by direct analyses of a major late mRNA species. A decapping-recapping method was used to specifically substitute a radioactively labeled phosphate for an unlabeled one within the cap structure. RNase H-susceptible sites were made by hybridizing synthetic oligodeoxyribonucleotides to the mRNA encoding a late major structural protein of 11 kilodaltons. Sequences of the type m7G(5')pppAmp (Ap)nUpG. . ., where n varies from a few to more than 40 nucleotides, were deduced by analysis of the length and sequence of RNase H, RNase T1, and RNase U2 digestion products.
通过对一种主要晚期mRNA种类进行直接分析，获得了位于翻译起始密码子紧邻上游的可变长度带帽多聚腺苷酸前导序列的证据。采用脱帽-重新加帽方法，在帽结构内用放射性标记的磷酸特异性替代未标记的磷酸。通过将合成的寡聚脱氧核糖核苷酸与编码11千道尔顿晚期主要结构蛋白的mRNA杂交，产生了核糖核酸酶H敏感位点。通过分析核糖核酸酶H、核糖核酸酶T1和核糖核酸酶U2消化产物的长度和序列，推导得出了m7G(5')pppAmp (Ap)nUpG... 类型的序列，其中n从几个核苷酸到40多个核苷酸不等。
