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鸭坦布苏病毒NS3蛋白的原核表达及其多克隆抗血清的制备 被引量:2
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作者 林霄 贺煜 +15 位作者 程安春 汪铭书 贾仁勇 朱德康 刘马峰 杨乔 吴英 赵新新 张沙秋 黄娟 刘韵雅 张玲 潘磊昌 欧旭敏 毛塞 陈舜 《四川农业大学学报》 CSCD 北大核心 2020年第5期619-623,共5页
【目的】利用原核表达系统表达鸭坦布苏病毒(Duck Tembusu virus,DTMUV)的NS3蛋白,并以此制备鼠抗NS3蛋白的多克隆抗血清。【方法】利用PCR扩增鸭坦布苏病毒CQW1株NS3基因片段,然后将其克隆到pET-28a(+)载体,构建重组质粒pET-28a(+)-NS... 【目的】利用原核表达系统表达鸭坦布苏病毒(Duck Tembusu virus,DTMUV)的NS3蛋白,并以此制备鼠抗NS3蛋白的多克隆抗血清。【方法】利用PCR扩增鸭坦布苏病毒CQW1株NS3基因片段,然后将其克隆到pET-28a(+)载体,构建重组质粒pET-28a(+)-NS3。将重组表达质粒转化到大肠杆菌BL21感受态细胞中,筛选获得阳性重组菌,经IPTG诱导表达,通过SDS-PAGE和Western blot试验表明重组NS3蛋白在BL21细胞中成功表达,将重组NS3蛋白切胶纯化后免疫小鼠后收获抗血清。【结果】经间接免疫荧光试验和Western blot试验验证,多克隆抗血清可识别病毒感染细胞过程产生的NS3蛋白和真核表达的NS3蛋白,具有良好的反应性和特异性。【结论】成功制备的DTMUV NS3蛋白的鼠源多克隆血清,将为鸭坦布苏病毒的临床检测和NS3蛋白功能性研究奠定基础。 展开更多
关键词 鸭坦布苏病毒 NS3蛋白 原核表达 多抗血清
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Alphaherpesvirus-vectored vaccines against animal diseases: Current progress 被引量:2
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作者 HU Yang WANG Ming-shu +24 位作者 CHENG An-chun JIA Ren-yong YANG Qiao WU Ying LIU Ma-feng ZHAO Xin-xin ZHU De-kang CHEN Shun ZHANG Sha-qiu WANG Yin GAO Qun OU Xu-min mao sai WEN Xing-jian XU Zhi-wen CHEN Zheng-li ZHU Ling LUO Qi-hui TIAN Bin PAN Lei-chang Mujeeb Ur REHMAN LIU Yun-ya YU Yan-ling ZHANG Ling CHEN Xiao-yue 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2020年第8期1928-1940,共13页
Recombinant virus-vectored vaccines are novel agents that can effectively activate specific and nonspecific immunity,are multivalent and multieffective,and have high safety ratings.Animal alphaherpesviruses have a lar... Recombinant virus-vectored vaccines are novel agents that can effectively activate specific and nonspecific immunity,are multivalent and multieffective,and have high safety ratings.Animal alphaherpesviruses have a large genome,contain multiple nonessential regions that do not affect viral replication and are capable of accepting the insertion of an exogenous gene and expressing the antigen protein.Furthermore,animal alphaherpesviruses have a wide host spectrum,can replicate in the host and continuously stimulate the animal to produce immunity to the corresponding pathogen,thus making them ideal carriers for recombinant virus-vectored vaccines.With the development of gene-editing technology,recombinant viruses capable of expressing foreign genes can be constructed by various methods.Currently,studies on recombinant virusvectored vaccines constructed based on animal alphaherpesviruses have involved poultry,pigs,cattle,sheep,and companion animals.Studies have shown that the construction of recombinant animal alphaherpesviruses enables the acquisition of immunity to multiple diseases.This article mainly summarizes the current progress on animal alphaherpesvirus-vectored vaccines,aiming to provide reference for the development of new animal alphaherpesvirus-vectored vaccines. 展开更多
关键词 recombinant vectored-vaccine recombinant alphaherpesviruses IMMUNITY gene-editing exogenous gene
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