摘要
本研究拟筛选出一组水疱性口炎病毒(VSV)拯救的最佳质粒转染比例,以提高病毒拯救效率。首先通过8组不同的转染比例进行病毒拯救,发现有5种比例能够成功拯救病毒,然后通过空斑分析病毒滴度,RT-q PCR检测病毒VSV-N m RNA的相对表达量,Western-blot分析VSV-G的表达。结果显示,与其他转染比例组相比,转染比例g组(pN∶pP∶pL∶VSV=3∶5∶1∶5)的拯救效率最优,转染比例b组(pN∶pP∶pL∶VSV=0.5∶0.4∶0.2∶1)的拯救效率最低。上述结果表明,本研究首次筛选出一组VSV病毒拯救的最佳质粒转染比例,为VSV反向遗传学的相关应用或研究提供了参考。
In order to increase the efficiency of virus rescue,our study carried out to screen the best transfection ratio for rescuing the vesicular stomatitis virus.First,virus rescue was conducted from 8 groups of various transfection ratios,among them,we verified that 5 groups of ratios could be successfully rescued.Plaque analysis was used to determine the virus titer,RT-qPCR was used to determine the relative mRNA expression of the virus VSV-N,and Western-blot was used to determine the expression of the VSV-G.Our results revealed that transfection ratio g(pN∶pP∶pL∶VSV=3∶5∶1∶5)had the highest rescue efficiency,whereas transfection ratio b(pN∶pP∶pL∶VSV=0.5∶0.4∶0.2∶1)was the lowest one.Taken together,our study is the first to evaluate the ideal plasmid transfection ratio for VSV rescue,providing fundamental for the application or study of VSV reverse genetics in the future.
作者
张永芝
杨帆
谭金龙
韩武玮怡
杨建社
张晋宽
刘瑞菊
赵伟杰
韩伟
何晓波
付宝权
李维克
ZHANG Yong-zhi;YANG Fan;TAN Jin-long;HAN Wu-wei-yi;YANG Jian-she;ZHANG Jin-kuan;LIU Rui-ju;ZHAO Wei-jie;HAN Wei;HE Xiao-bo;FU Bao-quan;LI Wei-ke(State Key Laboratory for Animal Disease Control and Prevention/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Hebei Key Laboratory of Preventive Veterinary Medicine/College of Animal Science and Technology,Hebei Normal University of Science&Technology,Qinhuangdao 066004,China;Northwest Agricultural and Forestry University/College of Life Sciences,Yangling 712100,China;Shengcheng Street Animal Husbandry and Veterinary Station of Shouguang City,Shouguang 262700,China;Anqiu Animal Husbandry Development Center,Anqiu 262100,China;Shandong Zhuohua Biotechnology Co.,Ltd,Liaocheng 252000,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2023年第9期1138-1144,共7页
Chinese Veterinary Science
基金
甘肃省科技重大专项(22ZD6NA001)
中国农业科学院兰州兽医研究所基本科研业务费项目(1610312021004)。
关键词
水疱性口炎病毒
病毒拯救
转染比例
vesicular stomatitis virus
virus rescue
transfection ratio
