Devos R, van Emmelo J, Seurinck-Opsomer C, Gillis E, Fiers W
Biochim Biophys Acta. 1976 Oct 18;447(3):319-27. doi: 10.1016/0005-2787(76)90055-1.
An oligo(A) or poly(A) segment was added in a stepwise fashion to the 3'-end of bacteriophage Qbeta-RNA with the aid of ATP : RNA adenylyltransferase from Escherichia coli. Nearly all RNA molecules, present in the reaction mixture, could be polyadenylated. For tail lengths not exceeding 200 nucleotide residues, the physical properties of Qbeta-RNA-poly(A) were found to be only slightly different from those of the original RNA. The polyadenylated RNA was purifed by affinity chromatography. The properties of Qbeta-RNA with oligo(A) tails of different average lengths were investigated in the in vitro replication reaction. Almost complete abolishment of template activity, even by short oligo(A) stretches, was found. Furthermore, polyadenylated Qbeta-RNA inhibited the normal replication reaction of Qbeta-RNA by removal of host factor HFI, in the same way as does free poly(A).
在来自大肠杆菌的ATP：RNA腺苷酸转移酶的帮助下，以逐步的方式将寡聚（A）或多聚（A）片段添加到噬菌体Qβ-RNA的3'末端。反应混合物中几乎所有的RNA分子都可以被聚腺苷酸化。对于长度不超过200个核苷酸残基的尾巴，发现Qβ-RNA-聚（A）的物理性质与原始RNA的物理性质仅略有不同。通过亲和层析纯化聚腺苷酸化的RNA。在体外复制反应中研究了具有不同平均长度寡聚（A）尾巴的Qβ-RNA的性质。发现即使是短的寡聚（A）片段也几乎完全消除了模板活性。此外，聚腺苷酸化的Qβ-RNA通过去除宿主因子HFI抑制Qβ-RNA的正常复制反应，其方式与游离聚（A）相同。
