Weber H
Biochim Biophys Acta. 1976 Jan 19;418(2):175-83. doi: 10.1016/0005-2787(76)90067-8.
The site of interaction of phage Qbeta coat protein with Qbeta RNA was determined by ribonuclease T1 degradation of complexes of coat protein and [32P]-RNA obtained by codialysis of the components from urea into buffer solutions. The degraded complexes were recovered by filtration through nitrocellulose filters, and bound [32P]RNA fragments were extracted and separated by polyacrylamide gel electrophoresis. Fingerprinting and further sequence analysis established that the three main fragments obtained (chain lengths 88, 71 and 27 nucleotides) all consist of sequences extending from the intercistronic region to the beginning of the replicase cistron. These results suggest that in the replication of Qbeta, as in the case of R17, coat protein acts as a translational repressor by binding to the ribosomal initiation site of the replicase cistron.
通过核糖核酸酶T1对噬菌体Qβ外壳蛋白与Qβ RNA复合物的降解来确定外壳蛋白与Qβ RNA的相互作用位点。该复合物是通过将各组分从尿素中透析到缓冲溶液中获得的。通过硝酸纤维素滤膜过滤回收降解后的复合物，提取结合的[32P]RNA片段，并通过聚丙烯酰胺凝胶电泳进行分离。指纹图谱和进一步的序列分析表明，获得的三个主要片段（链长分别为88、71和27个核苷酸）均由从基因间区域延伸至复制酶顺反子起始处的序列组成。这些结果表明，在Qβ的复制过程中，如同R17的情况一样，外壳蛋白通过与复制酶顺反子的核糖体起始位点结合而作为翻译阻遏物发挥作用。
