Mechanism of RNA-RNA Recombination in Brome Mosaic Virus: The Role of Viral Replicase

本研究的长期目标是了解溴代花叶病毒（BMV） RNA-RNA重组的分子机制，这是一种单链正义RNA病毒。我们在实验室开发了一种独特的异双工驱动重组系统，该系统能够指导突变体BMV RNA-3的工程3'非编码区与野生型病毒之间的杂交。我们将利用这个重组系统来研究BMV复制酶蛋白在重组中的作用。将确定1a和2a蛋白突变对交叉频率和对称性以及非模板核苷酸结合的影响。这些研究将使我们能够绘制重组活性复制酶序列。这些实验将为理解BMV和其他RNA病毒中重组事件的基本性质提供新的见解。* * *

9305389 Bujarski The long term goal of this research is to understand the molecular mechanism of RNA-RNA recombination in brome mosaic virus (BMV), a single-stranded positive-sense RNA virus. A unique heteroduplex driven recombination system capable of directing a cross between engineered 3' non-coding region of the mutant BMV RNA-3 and the wild type virus has been developed in our laboratory. We will use this recombination system to study the involvement of BMV replicase proteins in recombination. Effects of mutations in 1a and 2a proteins on the frequency and symmetry of crossovers and on the incorporation of nontemplated nucleotides will be determined. These studies should allow us to map the recombinationally active replicase sequences. %%% These experiments will provide novel insight into an understanding of the basic nature of recombinational events in BMV and other RNA viruses. ***