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人副流感病毒2型RNA在感染细胞中的特征及体外合成研究

Characterization of human parainfluenza virus type 2 RNAs in infected cells and by in vitro synthesis.

作者信息

Huang Y T, Romito R R, Panin M

机构信息

Institute of Pathology, Case Western Reserve University, Cleveland, Ohio 44106-4907, USA.

出版信息

Virus Res. 1995 Feb;35(2):181-92. doi: 10.1016/0168-1702(94)00095-t.

Abstract

The RNA species synthesized in HPIV-2 infected CV-1 cells were identified and characterized. The largest RNA of approximately 5.5 x 10(6) in molecular weight (MW) based on electrophoretic mobility, was identified as the genomic RNA. The other small RNA species of MWs 2.4 x 10(6), 1.1 x 10(6), 0.77 x 10(6), 0.68 x 10(6) and 0.5 x 10(6) were identified as mRNAs. The five smallest RNAs were also synthesized in vitro and comigrated with RNAs synthesized in virus-infected cells. mRNAs synthesized both in vitro and in virus-infected cells were translated in vitro. NP, P, M and V proteins synthesized in vitro comigrated, when analyzed by SDS-PAGE, with the authentic proteins synthesized in virus-infected cells. Additionally, peptide mapping showed that the NP, P and M proteins synthesized in vitro were indistinguishable from their counterparts synthesized in infected cells. Analysis of the proteins from virions identified L, HN, NP, F (F1, F2), P, M and V proteins as virion structural proteins. Electrophoretic mobility of reduced and nonreduced F proteins was found to be different due to the conformational changes conferred by disulfide bonds.

摘要

对在人副流感病毒2型(HPIV - 2)感染的CV - 1细胞中合成的RNA种类进行了鉴定和表征。根据电泳迁移率,分子量(MW)约为5.5×10⁶的最大RNA被鉴定为基因组RNA。分子量分别为2.4×10⁶、1.1×10⁶、0.77×10⁶、0.68×10⁶和0.5×10⁶的其他小RNA种类被鉴定为信使核糖核酸(mRNA)。这五种最小的RNA也在体外合成,并与在病毒感染细胞中合成的RNA共迁移。在体外和病毒感染细胞中合成的mRNA都在体外进行了翻译。当通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)分析时,体外合成的核蛋白(NP)、磷蛋白(P)、基质蛋白(M)和V蛋白与在病毒感染细胞中合成的相应天然蛋白共迁移。此外,肽图谱分析表明,体外合成的NP、P和M蛋白与在感染细胞中合成的对应蛋白无法区分。对病毒粒子中的蛋白质分析确定,L、血凝素 - 神经氨酸酶(HN)、NP、融合蛋白(F,F1、F2)、P、M和V蛋白为病毒粒子结构蛋白。由于二硫键赋予的构象变化,发现还原型和非还原型F蛋白的电泳迁移率不同。

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