Sugiura M, Miura K
Eur J Biochem. 1977 Feb 15;73(1):179-84. doi: 10.1111/j.1432-1033.1977.tb11305.x.
Double-stranded RNA of some virus genomes can be used as template for the DNA-dependent RNA polymerase purified from Escherichia coli. The RNA synthesis requires all four nucleoside triphosphates and manganese ions and is dependent on the presence of sigma subunit. The reaction is inhibited by rifampicin, streptolydigin and ethidium bromide, but not by DNase and actinomycin D which does not bind to double-stranded RNA. The template activity of double-stranded RNA from various viruses is different in each case. The order of template efficiency is Penicillum chrysogenum virus greater than cytoplasmic polyhedrosis virus greater than rice dwarf virus greater than reovirus. The product obtained using cytoplasmic polyhedrosis virus double-stranded RNA as template is single-stranded and hybridizes specifically to the denatured template RNA. One of the major 5'-starting nucleotide sequences of the product RNA is pppA-A-Y--. These results indicate that transcription in vitro of double-stranded RNA by E. Coli RNA polymerase is initiated at specific sites on the template.
某些病毒基因组的双链RNA可作为从大肠杆菌中纯化的依赖DNA的RNA聚合酶的模板。RNA合成需要所有四种核苷三磷酸和锰离子，并且依赖于σ亚基的存在。该反应受到利福平、链霉溶菌素和溴化乙锭的抑制，但不受DNase和不与双链RNA结合的放线菌素D的抑制。来自各种病毒的双链RNA的模板活性在每种情况下都不同。模板效率的顺序是产黄青霉病毒大于细胞质多角体病毒大于水稻矮缩病毒大于呼肠孤病毒。以细胞质多角体病毒双链RNA为模板获得的产物是单链的，并且与变性的模板RNA特异性杂交。产物RNA的主要5'-起始核苷酸序列之一是pppA-A-Y--。这些结果表明，大肠杆菌RNA聚合酶对双链RNA的体外转录是在模板上的特定位点起始的。
