RNA-RNA RECOMBINATION IN SINDBIS VIRUS

已知RNA病毒在自然界中迅速进化。新病毒的出现 毒力和组织及宿主特异性改变的菌株是一种 病毒进化的重要结果。RNA重组， 病毒进化中的关键力量导致基因组重排， 嵌合RNA基因组的形成。辛德比斯病毒（SIN）是一种蚊子 传播人类RNA病毒。包括风疹在内的许多脑炎病毒 与辛德毕斯病毒有关。先前已经证明， 两种转染到培养的动物细胞中的SIN RNA可以 经历重组，导致活病毒的产生。为继续 本研究中，序列和结构的要求， 将使用工程化RNA转录物探索参与的模板。 本研究旨在回答以下问题：1）什么是 3'和5'非病毒序列在重组中的作用？2)是什么 重组RNA交叉位点的序列。3)有没有 交叉位点的序列异质性？回答这些 问题，RNA模板与适当的修改将是 由相应的质粒合成。这些RNA可以 在体内进行重组，并通过以下方法分析子代RNA： 电泳，PCR介导的cDNA测序，和分析， 产生的序列。预计这些研究将有助于开发 基因工程RNA病毒疫苗。

RNA viruses are known to evolve rapidly in nature. Emergence of new virus strains with altered virulence and tissue and host specificity is an important consequence of virus evolution. RNA recombination, one of the critical forces in virus evolution results in genome rearrangements and formation of chimeric RNA genomes. Sindbis virus (SIN) is a mosquito transmitted human RNA virus. Many encephalitis viruses including Rubella virus are related to Sindbis virus. It was previously demonstrated that two SIN RNAs that were transfected into animal cells in culture can undergo recombination leading to production of live virus. In continuation of this study, the sequence and structural requirements of the participating templates will be explored using engineered RNA transcripts. This study specifically aims to answer the following questions: 1) What is the role of 3' and 5' non-viral sequences in recombination? 2) What are the sequences at the cross-over sites of recombinant RNAs. 3) Is there a sequence heterogeneity at the site of cross-over? To answer these questions, RNA templates with appropriate modifications will be synthesized from corresponding plasmids. These RNAs will be allowed to undergo recombination in vivo, and the progeny RNAs will be analyzed by electrophoresis, PCR mediated sequencing of cDNAs, and analysis of the resulting sequences. These studies are expected to aid in the development of genetically engineered RNA viral vaccines.