期刊论文

中文

中国兽医科学

1673-4696

2022

52

2

205-213

10.16656/j.issn.1673-4696.2022.0021

湖南省自然科学基金面上项目（2018JJ2177）； 湖南农业大学“双一流”建设项目（SYL2019048）；

本校为第一机构

为在昆虫细胞中表达猪流行性腹泻病毒（PEDV）的S和M蛋白，并探讨S和M蛋白能否自动组装成病毒样颗粒（VLPs）分泌至培养基中，本研究构建了3种不同的重组质粒（pFastBac-S、pFastBac-M和pFastBac-S-M），转化入DH10Bac感受态细胞，经蓝白斑筛选及PCR鉴定获得重组Bacmid，通过转染昆虫细胞Sf9获得重组杆状病毒（BV-S、BV-M 和BV-S-M）。在悬浮培养的昆虫细胞Sf9中进行表达，表达分泌的上清通过蔗糖梯度离心进行浓缩纯化，最终获得分泌型的S-M VLPs。通过Western-blotting分析S和M蛋白的表达和分泌特性，透射电镜观察VLPs形态，间接ELISA检测VLPs与猪PEDV阳性血清的反应性，最后免疫小鼠后检验其免...

In order to express S and M protein of porcine epidemic diarrhea virus in insect cells,and to explore whether the S and M protein could assemble automatically into VLPs and secrete into culture medium.In this work,three recombinant plasmids(p Fast Bac-S,p Fast Bac-M and p Fast Bac-S-M)were constructed,then the plasmids were transformed into competent cells DH10 Bac,and positive Bacmid DNA was screened through the blue-white screening and verified by PCR.The recombinant baculovirus(BV-S,BV-M and BV-S-M)were obtained by transfected insect cells S...