Brookes S M, Sun H, Dixon L K, Parkhouse R M
Institute for Animal Health, Pirbright Laboratory, Woking, Surrey, UK.
J Gen Virol. 1998 May;79 ( Pt 5):1179-88. doi: 10.1099/0022-1317-79-5-1179.
The j5R open reading frame (ORF) of the Malawi LIL 20/1 African swine fever virus (ASFV) isolate encodes a 111 amino acid protein with a putative transmembrane domain at the N terminus. Antisera raised against the predicted C-terminal peptide were used to identify the j5R protein by Western blotting in cells infected with ASFV or with recombinant vaccinia virus expressing the j5R ORF. This showed that the j5R protein migrates with an apparent molecular mass of 23-25 kDa, depending on the virus isolate, on SDS-PAGE and is expressed late during ASFV infection. The localization in infected cells and in virions of the j5R protein, and that of a previously characterized virion protein, j13L, which also contains a putative transmembrane domain, were studied by immunofluorescence and immuno-electron microscopy. Both proteins were expressed at 8-10 h post-infection (p.i.) as small multiple perinuclear foci which coalesced to a single area indicative of the virus factory at 18 h p.i. At the ultrastructural level j5R and j13L were detected mainly on membrane-like structures within the virus factory and on virus particles, suggesting that they may be involved in particle assembly. Negative contrast immuno-electron microscopy of mature extracellular virions confirmed that they are also integral structural proteins.
马拉维LIL 20/1非洲猪瘟病毒（ASFV）分离株的j5R开放阅读框（ORF）编码一种111个氨基酸的蛋白质，其N端有一个推定的跨膜结构域。用针对预测的C端肽产生的抗血清，通过蛋白质免疫印迹法在感染ASFV或表达j5R ORF的重组痘苗病毒的细胞中鉴定j5R蛋白。这表明，根据病毒分离株的不同，j5R蛋白在SDS-PAGE上的表观分子量为23 - 25 kDa，并且在ASFV感染后期表达。通过免疫荧光和免疫电子显微镜研究了j5R蛋白以及先前鉴定的病毒体蛋白j13L（其也包含一个推定的跨膜结构域）在感染细胞和病毒体中的定位。两种蛋白在感染后8 - 10小时（p.i.）均表达为多个小核周病灶，在感染后18小时合并为一个指示病毒工厂的单一区域。在超微结构水平上，j5R和j13L主要在病毒工厂内的膜样结构和病毒颗粒上被检测到，这表明它们可能参与颗粒组装。成熟细胞外病毒体的负染免疫电子显微镜证实它们也是完整的结构蛋白。
