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为筛选一些能抑制病毒RNA合成而不影响细胞DNA功能的抗病毒剂,在试管系统内,使用水稻普通矮缩病毒核糖核酸(RDV—RNA)转录酶和大肠杆菌(E·Coli·)依赖DNA的RNA聚合酶,在黑暗与照明条件下测试了若干染料与毒剂对这两种酶的活性的抑制作用。吖啶橙、溴化乙锭、美兰、天兰B(azureB)、天兰A(azureA)、天兰C(azureC)、劳斯氏紫(thionine)在P/D比率[RDV—RNA中的磷(P)对染料(D)的分子比率)为10/2及10/4时,抑制大肠杆菌DNA转录作用及RDV—RNA转录作用为40—70％,几乎是相同的。而氯奎、阿的平、藤黄醌茜素(luteoskyrin)、放线菌素D及利福霉素SV则仅抑制大肠杆菌DNA的转录作用,但不抑制RDV—RNA的转录作用。因为,已知属于第一类的某些染料能催化DNA与RNA的光动态反应,所以试验了天兰B对RDV—RNA同一反应的催化效果。实际上,当反应混合物以20000勒克司照明,夭兰B在P/D比率低达10/0.06时,抑制RDV—RNA转录作用约为50％。当RDV—RNA在有天兰B时照明,P/D比率在10/0.12以上时,仅鸟嘌呤受损失。当水稻普矮病毒颗粒在照明下用天兰B进行予处理,然后加入反应混合物,在D/D比率为10/0.015时照明30分钟后,RDV—RNA转录作用出现50％的抑制。用~(32)P标记的水稻普矮病毒(RDV)颗粒在有天兰B时照明,并用苯酚抽提RNA,在抽提的RNA中测不到鸟嘌呤的损失,但RNA的量下降,相反地,随着天兰B剂量的增加,~(32)P结合于旦白质的量增多。从这些结果可以断定,在黑暗中天兰B引起的抑制作用是由于染料嵌入到双链的RNA中所引起,但在照明下发生的抑制则是由于受染料催化的光动态反应所引起,而且这种抑制比发生于黑暗的更加有效。因为白天植株暴露于阳光下,所以后一机理在用天兰B处理感染了水稻普矮病毒的稻株中可能占优势。
To screen for some antiviral agents that inhibit the synthesis of viral RNA without affecting the DNA function of cells, the RDV-RNA transcriptase and E.coli-dependent DNA RNA polymerase was used to test the inhibitory effect of several dyes and poisons on the activity of both enzymes under dark and light conditions. Acridine orange, ethidium bromide, melanopterin, azureB, azureA, azureC, and thionine in the P / D ratio [RDV-RNA Of phosphorus (P) to dye (D)) was 10/2 and 10/4, the inhibition of DNA transcription in E. coli and RDV-RNA transcription was 40-70% and almost the same. However, chloroquine, azepin, luteoskyrin, actinomycin D and rifamycin SV inhibited the transcription of E. coli DNA only, but did not inhibit the transcription of RDV-RNA. Because it is known that some dyes belonging to the first class catalyze the photodynamic reaction of DNA with RNA, the catalytic effect of Celastrus B on the same reaction of RDV-RNA was tested. In fact, when the reaction mixture was illuminated at 20000 lux, ylanglanb inhibited RDV-RNA transcription by about 50% at P / D ratios as low as 10 / 0.06. When RDV-RNA was illuminated with Celasanil B, only guanine was lost when the P / D ratio was above 10 / 0.12. When rice PUFV particles were treated with Celastrobin B under illumination and then added to the reaction mixture, a 50% inhibition of RDV-RNA transcription was observed after 30 minutes of illumination at a D / D ratio of 10 / 0.015. The ~ (32) P-labeled RDV particles were illuminated when there was Celastrus B and the RNA was extracted with phenol. The loss of guanine was not detected in the extracted RNA, but the amount of RNA decreased. Conversely, as the dose of Celastrus B increased, the amount of ~ (32) P bound to the protein increased. From these results, it can be concluded that the inhibitory effect of Celastrus B in the dark is caused by the dye being embedded in the double-stranded RNA, but the inhibition due to illumination is due to the photodynamic reaction catalyzed by the dye, and This suppression is more effective than darkness. Because the plants were exposed to sunlight during the daytime, the latter mechanism may be predominant in the treatment of rice plants infected with the rice Puti virus with Celastrus B.