利用杆状病毒表达系统克隆及表达斑点叉尾鮰病毒假定糖蛋白基因

Cloning and expression of a putative glycoprotein gene of channel catfish virus using baculovirus expression system.

作者信息

Kucuktas H, Brady Y J, Tuzun S

机构信息

Department of Fisheries and Allied Aquacultures, Auburn University, Alabama 36849, USA.

出版信息

Dis Aquat Organ. 1998 Nov 30;34(3):231-7. doi: 10.3354/dao034231.

DOI:10.3354/dao034231

PMID:9891736

Abstract

The 1035 base-pair (bp) open reading frame 59 (ORF59) of channel catfish virus (CCV) codes for an abundant hydrophobic membrane glycoprotein considered to be the major envelope glycoprotein of the virus. The ORF59 gene of CCV was cloned into the baculovirus genome by site-specific transposition and expressed in Spodoptera frugiperda cells (Sf9 cells). Expressed protein could be recognized by polyclonal antibody against CCV and appeared to have the same electrophoretic mobility as native protein. Crude lysates of Sf9 cells infected with recombinant baculovirus containing ORF59 gene inhibited plaque formation of CCV on channel catfish ovary cells.

摘要

斑点叉尾鮰病毒（CCV）的1035个碱基对（bp）的开放阅读框59（ORF59）编码一种丰富的疏水膜糖蛋白，该蛋白被认为是该病毒的主要包膜糖蛋白。通过位点特异性转座将CCV的ORF59基因克隆到杆状病毒基因组中，并在草地贪夜蛾细胞（Sf9细胞）中表达。表达的蛋白能被抗CCV的多克隆抗体识别，并且其电泳迁移率似乎与天然蛋白相同。用含有ORF59基因的重组杆状病毒感染的Sf9细胞的粗裂解物可抑制CCV在斑点叉尾鮰卵巢细胞上形成蚀斑。