关键词: 蜱传森林脑炎病毒, 非结构蛋白1, 原核表达, 融合蛋白

Abstract: Abstract:Objective To construct the prokaryotic expression vector of  nonstructural protein 1(NS1) of Tick-borne encephalitis virus(TBEV) and to o
btain  high purity expression of NS1 and to provide  experimental basis for preparation of polyclonal antibody against NS1 and study on its function.
Methods The PCR primers were designed according to the target gene,and the prokaryotic expression vector pEASYTM-E1-NS1 was constructed using gene recombination method.The recombinant plasmids pEASYTM-E1-NS1 were transferred into BL21 cells,and the expressed fusi
on proteins were purified by nickel ion affinity purification and  the concentration of protein was analyzed by Bradford method after indu
ction by Isopropy1β-D-1-Thiogalactopyranoside(IPTG).The expression level of fusion protein was detected by SDS-PAGE
and Western blotting methods.Results The prokaryotic expression vector pEASYTM-E1-NS1 carrying  TBEV NS1  expressed fusion protein in the form of
inclusion body after induction;a kind of highly purified protein was obtained through affinity purification. The SDS-PAGE and Western blotting results showed that a kind of specific protein with a relative molecular mass of about 40 000 was found.Conclusion The prokaryotic expression vector pEASYTM-E1-NS1 is constructed successfully,and the fusion protein of  TBEV NS1 with  high purity  is acquired.

Key words: Tick-borne encephalitis virus, nonstructural protein 1, prokaryotic expression, fusion protein