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莴苣传染性黄化病毒:使用部分纯化的病毒粒子和烟粉虱进行体外获毒分析

Lettuce infectious yellows virus: in vitro acquisition analysis using partially purified virions and the whitefly Bemisia tabaci.

作者信息

Tian T, Rubio L, Yeh H H, Crawford B, Falk B W

出版信息

J Gen Virol. 1999 May;80 ( Pt 5):1111-1117. doi: 10.1099/0022-1317-80-5-1111.

Abstract

Virions of lettuce infectious yellows virus (LIYV; genus Crinivirus) were purified from LIYV-infected plants and their protein composition was analysed by SDS-PAGE and immunoblotting. Virion preparations contained the major capsid protein (CP), but the minor capsid protein (CPm), p59 and the HSP70 homologue were also identified by immunoblot analysis. Immunogold labelling analysis showed that CP constituted the majority of the LIYV virion capsid, but CPm was also part of the capsid and localized to one end of the virion, similar to the polar morphology seen for viruses in the genus Closterovirus. p59 and the HSP70 homologue were not detected on virions by immunogold labelling, but were always detected in virion preparations by immunoblot analysis. Purified LIYV virions were used for in vitro acquisition analysis with Bemisia tabaci whiteflies and were efficiently transmitted to plants. Infectivity neutralization analyses were done using antisera to the LIYV-encoded CP, CPm, p59 and HSP70 homologue. Only antiserum to the CPm effectively neutralized LIYV transmission by B. tabaci. These data suggest that the LIYV-B. tabaci transmission determinants are associated with purified virions, and that the LIYV virion structural protein CPm is involved in transmission by B. tobaci.

摘要

从感染生菜传染性黄化病毒(LIYV;毛形病毒属)的植物中纯化出病毒粒子,并通过SDS-PAGE和免疫印迹分析其蛋白质组成。病毒粒子制剂含有主要衣壳蛋白(CP),但通过免疫印迹分析也鉴定出了次要衣壳蛋白(CPm)、p59和HSP70同源物。免疫金标记分析表明,CP构成了LIYV病毒粒子衣壳的大部分,但CPm也是衣壳的一部分,并定位于病毒粒子的一端,类似于在黄症病毒属病毒中看到的极性形态。通过免疫金标记在病毒粒子上未检测到p59和HSP70同源物,但通过免疫印迹分析在病毒粒子制剂中总能检测到它们。纯化的LIYV病毒粒子用于与烟粉虱进行体外获毒分析,并有效地传播到植物中。使用针对LIYV编码的CP、CPm、p59和HSP70同源物的抗血清进行感染性中和分析。只有针对CPm的抗血清有效地中和了烟粉虱传播LIYV的能力。这些数据表明,LIYV-烟粉虱传播决定因素与纯化的病毒粒子有关,并且LIYV病毒粒子结构蛋白CPm参与了烟粉虱的传播。

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