McMillan D J, Kay J, Mills J S
Department of Biology, Roche Discovery Welwyn, UK.
J Gen Virol. 1997 Sep;78 ( Pt 9):2153-7. doi: 10.1099/0022-1317-78-9-2153.
Varicella-zoster virus (VZV) genes 33 and 33.5 are predicted to encode the VZV proteinase and its substrate (the assembly protein) respectively. These genes were expressed in insect cells using recombinant baculovirus and it was confirmed that gene 33 encodes a proteinase capable of autoproteolytic processing at two positions. When VZV gene 33.5 was co-expressed with the VZV proteinase, processing of the VZV33.5 gene product was observed. A polyclonal antiserum to the VZV assembly protein domain highlighted a set of proteins in VZV infected HEL cells identical to those identified in insect cells expressing VZV genes 33 and 33.5. To facilitate further characterization of the VZV proteinase the enzyme was purified by affinity chromatography from an E. coli expression system and in vitro activity was observed.
水痘带状疱疹病毒(VZV)的基因33和33.5预计分别编码VZV蛋白酶及其底物(装配蛋白)。利用重组杆状病毒在昆虫细胞中表达了这些基因,并且证实基因33编码一种能够在两个位点进行自催化加工的蛋白酶。当VZV基因33.5与VZV蛋白酶共表达时,观察到了VZV33.5基因产物的加工过程。针对VZV装配蛋白结构域的多克隆抗血清突出显示了VZV感染的HEL细胞中的一组蛋白质,这些蛋白质与在表达VZV基因33和33.5的昆虫细胞中鉴定出的蛋白质相同。为了便于对VZV蛋白酶进行进一步表征,通过亲和层析从大肠杆菌表达系统中纯化了该酶,并观察到了其体外活性。