使用CRISPR/Cas9技术构建新型重组伪狂犬病毒疫苗的初步研究  

PRELIMINARY STUDY ON UTILIZING CRISPR/CAS9 TO CONSTRUCT NEW RECOMBINANT PSEUDORABIES VARIANT VACCINE

作　　者：于之清[1] 童武[1] 郑浩[1] 李国新[1] 高飞[1] 王涛[1] 梁超[1] 叶超[1] 武吉强 黄勤峰 童光志[1] YU Zhi-qing TONG Wu ZHENG Hao LI Guo-xin GAO Fei WANG Tao LIANG Chao YE Chao WU Ji-qiang HUANG Qin-feng TONG Guang-zhi(Shanghai Veterinary Research Institute, CAAS, Shanghai 200241, China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2017年第4期6-12,共7页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点研发计划项目(2016YFD0500100)

摘　　要：本研究使用CRISPR/Cas9基因编辑技术结合同源重组,以伪狂犬病毒经典疫苗株Bartha-K61为骨架,将其gB基因替换为伪狂犬病毒变异株JS-2012的gB基因,经噬斑纯化、PCR筛选鉴定,获得重组病毒r PRV-BJB。体外生物学特性分析结果显示,重组病毒株r PRV-BJB与疫苗株Bartha-K61具有相似的生长特性和噬斑形态,可作为新型重组伪狂犬病病毒疫苗候选株。In this study, we utilized the specific CRISPR/Cas9 gene editing system combined with homologous recombination to replace the gB of Bartha-K61 strain with that of JS-2012 strain. After several rounds plaque purification and PCR-screening, the acquired recombinant virus was identified by sequencing and designated as r PRV-BJB. r PRV-BJB and Bartha-K61 showed similar biological characteristics in vitro, indicating that r PRV-BJB is a promising PRV vaccine candidate that can be performed in following protective efficacy evaluation.

关 键 词：变异伪狂犬病毒 CRISPR/Cas9 重组病毒 GB基因 

分 类 号：S852.659.1[农业科学—基础兽医学]