Cancedda R, Shatkin A J
Eur J Biochem. 1979 Feb 15;94(1):41-50. doi: 10.1111/j.1432-1033.1979.tb12869.x.
Sindbis virus 42-S and 26-S RNAs labeled with 32P were purified from infected chick embryo fibroblasts. The RNA's were incubated in the presence of a wheat germ cell-free translating system under conditions that yielded 40-S and 80-S initiation complexes. After digestion with RNase A, ribosome-protected fragments were isolated by polyacrylamide gel electrophoresis and compared with respect to number, size, cap content and oligonucleotide composition. The two RNA species yielded several fragments of chain length about 35--40 nucleotides from 80S complexes and up to 60--65 nucleotides from 40-S complexes. The 5'-terminal capped sequence, m7 GpppA-U-G that is present in both Sindbis virus RNA's, was not retained in any of the ribosome-protected fragments. Fingerprint analyses indicated that the fragments derived from 40S and 80-S initiation complexes of each species of RNA were overlapping, but the fragments from 42-S and 26-S RNAs were unrelated. The complexity of the fingerprints were consistent with protection of a single, different initiation site in each Sindbis virus RNA.
用32P标记的辛德毕斯病毒42-S和26-S RNA从感染的鸡胚成纤维细胞中纯化出来。将这些RNA在麦胚无细胞翻译系统存在的条件下孵育，该条件可产生40-S和80-S起始复合物。用核糖核酸酶A消化后，通过聚丙烯酰胺凝胶电泳分离核糖体保护片段，并在片段数量、大小、帽结构含量和寡核苷酸组成方面进行比较。这两种RNA种类从80S复合物产生了几个链长约35 - 40个核苷酸的片段，从40-S复合物产生了多达60 - 65个核苷酸的片段。辛德毕斯病毒两种RNA中都存在的5'-末端帽结构序列m7GpppA-U-G，在任何核糖体保护片段中都未保留。指纹分析表明，每种RNA的40S和80-S起始复合物衍生的片段相互重叠，但42-S和26-S RNA的片段不相关。指纹的复杂性与每种辛德毕斯病毒RNA中单个不同起始位点的保护一致。
