狂犬病病毒aG株假病毒的制备方法  

Preparation Method of Rabies Virus aG Strain Pseudovirus

作　　者：张雨薇 韩德明[1] ZHANG Yuwei;HAN Deming(School of Life Science and Technology,Changchun University of Science and Technology,Changchun 130022)

机构地区：[1]长春理工大学生命科学技术学院,长春130022

出　　处：《长春理工大学学报（自然科学版）》2023年第1期102-107,共6页Journal of Changchun University of Science and Technology(Natural Science Edition)

基　　金：吉林省科技发展计划项目(20200201099JC)。

摘　　要：基于HIV慢病毒包装系统建立了狂犬病病毒(RV)aG株假病毒的制备方法。构建编码RV aG株糖蛋白(GP)的重组真核表达质粒pCMV-aG-G,转染至293T细胞验证GP的表达。将GP表达质粒与假病毒包装质粒及绿色荧光蛋白(GFP)指示质粒共转染293T细胞进行假病毒包装,制备携带GFP报告基因与aG株GP的RV假病毒。结果表明,研究成功构建了编码RV aG株GP的重组真核表达质粒,其可在293T细胞中有较高表达;成功制备了具有细胞感染性的RV aG株假病毒。本研究有助于更安全、准确监测针对我国现用人狂犬病疫苗生产用RV固定毒aG株的中和抗体水平。The preparation method of rabies virus(RV) aG strain pseudovirus was established based on HIV lentiviral packaging system.The recombinant eukaryotic expression plasmid pCMV-aG-G encoding RV aG strain glycoprotein(GP) was constructed and transfected into 293T cells to verify the expression of GP.The GP expression plasmid,the pseudovirus packaging plasmid and the green fluorescent protein(GFP) indicator plasmid were co-transfected into 293T cells for pseudovirus packaging;and the RV pseudovirus carrying the GFP reporter gene and aG strain GP was prepared.The results showed that a recombinant eukaryotic expression plasmid encoding RV aG strain GP was successfully constructed in this study,which could be highly expressed in 293T cells;a cell-infectious RV aG strain pseudovirus was successfully prepared.This study contributes to safer and more accurate monitoring of neutralizing antibody levels against the immobilized aG strain of RV used in the production of human rabies vaccines in China.

关 键 词：狂犬病病毒 假病毒 aG株 转染 

分 类 号：R373.9[医药卫生—病原生物学]