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Vaccinia Virus Gene B7R Encodes an 18-kDa Protein That is Resident in the Endoplasmic Reticulum and Affects Virus Virulence

机译:牛痘病毒基因B7R编码内质网中存在的18 kDa蛋白并影响病毒毒力。

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摘要

This paper presents a characterisation of vaccinia virus (VV) gene B7R that was predicted to encode a polypeptide of 182 amino acids with an N-terminal signal peptide. In vitro transcription and translation analysis showed the B7R gene product was a 21-kDa protein that, in the presence of microsomes, was processed into an 18-kDa mature form. The 18-kDa form associated with the microsomal membranes and was within the lumen of the vesicle where it was inaccessible to exogenous protease or an antibody raised against the B7R C terminus. Within VV-infected cells, the 18-kDa form of B7R was detected late during infection in the endoplasmic reticulum where it colocalised with protein disulphide isomerase. The B7R protein was detected neither in the culture supernatant nor associated with virus particles. A virus deletion mutant lacking the B7R gene and a revertant virus were constructed. Compared to wild-type and revertant viruses, the deletion mutant replicated normally in cell culture and had unaltered virulence in a murine intranasal model of infection. However, the deletion mutant was attenuated in a murine intradermal model where it induced a smaller lesion than the control viruses.
机译:本文介绍了牛痘病毒(VV)基因B7R的特征,该基因预计可编码带有N端信号肽的182个氨基酸的多肽。体外转录和翻译分析表明B7R基因产物是一种21 kDa的蛋白质,在微粒体存在下,该蛋白质被加工成18 kDa的成熟形式。 18-kDa形式与微粒体膜有关,位于囊泡内腔,在此处无法接触外源蛋白酶或针对B7R C末端的抗体。在被VV感染的细胞中,在感染后期在内质网中检测到18kDa形式的B7R,并与蛋白质二硫键异构酶共定位。在培养上清液中既未检测到B7R蛋白,也未与病毒颗粒相关联。构建了缺少B7R基因的病毒缺失突变体和回复病毒。与野生型和回复型病毒相比,缺失突变体可在细胞培养物中正常复制,并且在鼠鼻内感染模型中具有不变的毒力。然而,该缺失突变体在鼠皮内模型中被减毒,在该模型中它引起的病变比对照病毒小。

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