中国动物传染病学报2025，Vol.33Issue(2)：139-145,7.
犬副流感病毒F基因原核表达及多克隆抗体的制备
Prokaryotic Expression of F Gene of Canine Parainfluenza Virus and Preparation of Polyclonal Antibodies
摘要
Abstract
In this study,a prokaryotic expression system was used to express Canine parainfluenza virus(CPIV)F protein for preparation of polyclonal antibodies.The primers were designed according to the F gene of CPIV SH2021 strain(registration number OL548910),and the F gene fragment of the signal peptide was amplified to construct the recombinant expression plasmid pET-42a-CPIV-F,which was then converted into E.coli BL21 after sequencing verification.The expression of the recombinant F protein was induced with optimal IPTG.The recombinant CPIV-F protein was purified by inclusion body crude purification for preparation of rabbit polyclonal antibodies.The results showed that the pET-42a-CPIV-F prokaryotic expression plasmid was constructed and the recombinant CPIV-F protein was expressed in E.coli BL21.The recombinant CPIV-F protein was visualized as a molecular mass of about 89 kDa in SDS-PAGE and Western blot.The optimized expression conditions included induction of IPTG at 1 mmol/L for 4 h,resulting in the recombinant protein concentration of 2.3 mg/mL.Rabbit anti-CPIV-F antibodies specifically reacted with the recombinant proteins in Western blot and showed a titer of 1∶102 400 in indirect ELISA.The rabbit polyclonal antibodies prepared in this study were specific and reactive,indicating that F protein had good immunogenicity.In summary,this study laid a foundation for further study of the biological function of CPIV F protein and development of immunological detection methods.关键词
犬副流感病毒/F基因/原核表达/多克隆抗体Key words
Canine parainfluenza virus/F gene/prokaryotic expression/polyclonal antibodies分类
农业科技引用本文复制引用
刘禹含,李传峰,叶雨濛,麦嘉迅,程松,孟春春,朱杰,刘光清,王勇,巴音查汗·盖力克..犬副流感病毒F基因原核表达及多克隆抗体的制备[J].中国动物传染病学报,2025,33(2):139-145,7.基金项目
国家自然科学基金项目(32000109) （32000109）
上海市青年科技英才扬帆计划项目(20YF1457700) （20YF1457700）
上海市自然科学基金项目(22ZR1476300) （22ZR1476300）
上海市市级科技重大专项(ZD2021CY001) （ZD2021CY001）
中央级公益性行业科院所基本科研业务费(2022JB01) （2022JB01）
