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棉铃虫5型质型多角体病毒属于呼肠孤病毒科质型多角体病毒属,以重要农业害虫棉铃虫为其天然宿主,对棉铃虫的生物控制具有重要意义.本文对棉铃虫5型质型多角体病毒第3片段编码的蛋白的功能进行了初步研究.首先通过同源性对比,推测其所编码的蛋白可能行使RNA依赖的RNA聚合酶(RdRP)的功能.通过体外活性研究确定了该蛋白的RdRP活性,并确定了其保守活性位点GDD.随后以病毒基因组RNA和3′-OH封闭的病毒基因组RNA为模板,利用Northern blot方法研究该蛋白起始病毒基因组RNA合成的分子机制.结果表明,该病毒的RdRP主要通过引物非依赖的方式起始病毒基因组RNA的合成,并且该RdRP蛋白并不具有末端转移酶活性.最后,对RdRP行使功能的生化条件进行探索,发现RdRP功能的发挥需要二价金属离子Mg2+的存在.
Helicoverpa armigera 5 type polyhedrosis virus belongs to the reovirus genus polyhedrosis genus, and the important agricultural pest Helicoverpa armigera is its natural host, which is of great significance to the biological control of the cotton bollworm.In this paper, Type polyhedrosis virus fragment encoded by the first part of the function of a preliminary study.First, through homology comparison, it is speculated that the encoded protein may exercise RNA-dependent RNA polymerase (RdRP) function by in vitro activity study identified The RdRP activity of this protein was determined and the GDD of its conserved active site was determined. The molecular mechanism of RNA synthesis of this protein was also investigated by Northern blot using viral genomic RNA and 3’-OH-enclosed viral genomic RNA as template The results showed that RdRP mainly initiated the synthesis of viral genomic RNA in a primer-independent manner, and the RdRP protein did not have terminal transferase activity.Finally, the biochemical conditions of RdRP function were explored and found that RdRP function The play requires the presence of divalent metal ions Mg2 +.