Samokhvalov E I, Iuferov V P, Uryvaev L V, Zhdanov V M
Vopr Virusol. 1983 May-Jun(3):270-3.
A method for isolation of terminal polyriboadenylate transferase from E. coli cells (MPE 600) is presented. The specific activity of the enzyme yield at the terminal stage was 933 units per 1 mg of protein. Analysis of polyadenylated in vitro virion RNA of influenza virus A/USSR/90/17 strain in polyacrylamideagarose gel (2.2%-0.6%) in the presence of 6 M urea showed all the 8 fragments of genome RNA to be adenylated, their sizes being retained with regard to the distribution in gel of the initial RNA fragments. In vitro polyadenylated virion RNA was an effective matrix in reverse transcription reaction with RNA-dependent DNA-polymerase using oligo (dT) as a primer. Complementary DNAs obtained in this way may be the starting material for synthesis of double-stranded DNAs and subsequent construction of recombinant DNAs containing influenza virus genetic information.
本文介绍了一种从大肠杆菌细胞（MPE 600）中分离末端聚核糖腺苷酸转移酶的方法。在末期阶段获得的该酶的比活性为每1毫克蛋白质933单位。在6 M尿素存在下，对甲型流感病毒A/苏联/90/17株的体外聚腺苷酸化病毒粒子RNA在聚丙烯酰胺 - 琼脂糖凝胶（2.2%-0.6%）中进行分析，结果表明基因组RNA的所有8个片段均被腺苷酸化，其大小相对于初始RNA片段在凝胶中的分布得以保留。体外聚腺苷酸化的病毒粒子RNA在用寡聚（dT）作为引物的依赖RNA的DNA聚合酶的逆转录反应中是一种有效的模板。以这种方式获得的互补DNA可以作为合成双链DNA以及随后构建包含流感病毒遗传信息的重组DNA的起始材料。
