首页> 美国卫生研究院文献>Journal of Virology >Identification Mapping and In Vitro Translation of Campoletis sonorensis Virus mRNAs from Parasitized Heliothis virescens Larvae
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Identification Mapping and In Vitro Translation of Campoletis sonorensis Virus mRNAs from Parasitized Heliothis virescens Larvae

机译:鉴定定位和体外翻译从寄生的Heliothis virescens幼虫的Campontis sonorensis病毒mRNAs。

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摘要

Expression of Campoletis sonorensis virus (CsV) in parasitized Heliothis virescens larvae was investigated by Northern blot analysis of poly(A)+ mRNAs isolated from H. virescens larvae at various times after parasitization by C. sonorensis. At least 12 CsV mRNAs were detected in parasitized H. virescens larvae. Injection of nonparasitized H. virescens larvae with purified CsV resulted in a pattern of viral mRNAs similar to that observed in naturally parasitized larvae. With CsV DNA restriction fragments which contained expressed sequences, individual CsV mRNAs were mapped to the superhelical DNAs of the viral genome. Two gene-specific probes, which consisted of cloned S1 nuclease-protected restriction fragments, each hybridized to several CsV superhelical DNAs, suggesting that some CsV genes may be shared on several superhelical DNAs. Cloned restriction fragments containing sequences which flank the expressed sequences also hybridized to numerous CsV superhelical DNAs. Some CsV proteins were identified by in vitro translation of hybrid-selected CsV mRNAs.
机译:通过对C. sonorensis进行寄生化处理后的不同时间,从分离自H. virescens幼虫的poly(A) + mRNA的Northern印迹分析,研究了Campontis sonorensis病毒(CsV)在被寄生的Heliothis virescens幼虫中的表达。在寄生的H. virescens幼虫中至少检测到12个CsV mRNA。用纯化的CsV注射未寄生虫的荚膜梭菌幼虫会产生与天然寄生虫的幼虫相似的病毒mRNA模式。使用包含表达序列的CsV DNA限制片段,将单个CsV mRNA定位到病毒基因组的超螺旋DNA。由克隆的S1核酸酶保护的限制性片段组成的两个基因特异性探针,每个都与几个CsV超螺旋DNA杂交,表明某些CsV基因可能在几个超螺旋DNA上共享。含有位于表达序列侧翼的序列的克隆的限制性片段也与许多CsV超螺旋DNA杂交。通过体外选择杂交的CsV mRNA鉴定了一些CsV蛋白。

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