CAJ | 학술논문

本文以嗜肾型鸡传染性支气管炎病毒（Infectious bronchitis virus,IBV）毒株的RNA为模板,通过RT-PCR扩增获得IBV非结构蛋白5（non-structural protein,nsp5）基因片段后,构建了杆状病毒重组质粒IBVnsp5-Bacmid。IBVnsp5-Bacmid转染Sf9细胞,获得nsp5重组杆状病毒,经（immunofluorescence assay,IFA）和Western blot检测到转染细胞表达的nsp5蛋白。进一步从感染的细胞中纯化重组蛋白,并用纯化的蛋白免疫小鼠制备了抗nsp5的多抗血清,该多抗血清可检测到IBV四川株SC021202感染的DF-1细胞中特异性的nsp5蛋白。结果表明IBV nsp5在Bac-to-Bac真核表达系统中获得了成功表达,而且具有良好的免疫原性和反应原性。
본문이기신형계전염성지기관염병독（Infectious bronchitis virus,IBV）독주적RNA위모판,통과RT-PCR확증획득IBV비결구단백5（non-structural protein,nsp5）기인편단후,구건료간상병독중조질립IBVnsp5-Bacmid。IBVnsp5-Bacmid전염Sf9세포,획득nsp5중조간상병독,경（immunofluorescence assay,IFA）화Western blot검측도전염세포표체적nsp5단백。진일보종감염적세포중순화중조단백,병용순화적단백면역소서제비료항nsp5적다항혈청,해다항혈청가검측도IBV사천주SC021202감염적DF-1세포중특이성적nsp5단백。결과표명IBV nsp5재Bac-to-Bac진핵표체계통중획득료성공표체,이차구유량호적면역원성화반응원성。
Little is known about the function of non-structural proteins 5（nsp5） of avian infectious bronchitis virus（IBV）.In this study,IBV nsp5 was amplified in RT-PCR using RNA of nephropathogenic IBV strain as template.The recombinant plasmid IBVnsp5-Bacmid was constructed based on Bac-to-Bac baculovirus expression system and transfected into Sf9 cells.The expression of recombinant nsp5 in Sf9 cells was confirmed by IFAT and Western blot analysis.The polyclonal antiserum against nsp5 was prepared using recombinant nsp5 purified from transfected Sf9 cells and demonstrated to react with IBV SC021202 infected DF-1 cells in IFAT.The results indicated that IBV nsp5 was successfully expressed in eukaryotic expression system and had good angenicity.