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斜纹夜蛾多粒包埋核型多角体病毒中一种与封闭型衍生病毒包膜相关的新型蛋白质的鉴定

Identification of a novel protein associated with envelope of occlusion-derived virus in Spodoptera litura multicapsid nucleopolyhedrovirus.

作者信息

Yin Chong, Yu Jianxiu, Wang Lihua, Li Zhaofei, Zhang Ping, Pang Yi

机构信息

State Key Laboratory for Biocontrol, Institute of Entomology, Zhongshan University, Guangzhou 510275, China.

出版信息

Virus Genes. 2003 Jan;26(1):5-13. doi: 10.1023/a:1022310202540.

Abstract

Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV) ORF137 (Splt137) is one of 29 unique SpltMNPV ORFs. Splt137 has the potential to code for a polypeptide of 231 amino acid residues with predicted molecular weight of 27.5 kDa. Computer-assisted analysis of the predicted amino acid sequences of Splt137 protein showed 1 N-glycosylation site and 11 phosphorylation sites. For identification of Spit137, antibody was prepared by immunization of rabbits with purified Splt137 protein produced in Escherichia coli. This antibody was used to analyse Splt137 protein using Western blot. A 36-kDa protein was found both in the infected cells and envelope fractions of occlusion-derived virus (ODV) but could not be detected in the budded virus (BV). Tunicamycin treatment of SpltMNPV infected cells suggested that the 36-kDa protein had undergone N-glycosylation. Our data suggested that Splt137 protein was a novel envelope protein of ODV and might exist as a more complex form of 79-kDa protein in intact ODV. Further, transcriptional analysis with RT-PCR and 5' RACE analysis suggested that Splt137 might perform functions early and late in infection.

摘要

斜纹夜蛾多粒包埋核多角体病毒(SpltMNPV)的开放阅读框137(Splt137)是SpltMNPV 29个独特的开放阅读框之一。Splt137有可能编码一个由231个氨基酸残基组成的多肽,预测分子量为27.5 kDa。对Splt137蛋白预测氨基酸序列的计算机辅助分析显示有1个N-糖基化位点和11个磷酸化位点。为了鉴定Splt137,用在大肠杆菌中产生的纯化Splt137蛋白免疫兔子制备了抗体。该抗体用于通过蛋白质免疫印迹法分析Splt137蛋白。在感染细胞和多角体衍生病毒(ODV)的包膜组分中均发现了一种36 kDa的蛋白,但在出芽病毒(BV)中未检测到。用衣霉素处理SpltMNPV感染的细胞表明,36 kDa的蛋白发生了N-糖基化。我们的数据表明,Splt137蛋白是ODV的一种新型包膜蛋白,在完整的ODV中可能以79 kDa蛋白的更复杂形式存在。此外,用逆转录-聚合酶链反应(RT-PCR)进行的转录分析和5′端快速扩增cDNA末端(5′RACE)分析表明,Splt137可能在感染的早期和晚期发挥作用。

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