收稿日期:2022-07-13基金项目:吉林省科技发展计划项目(20200201099JC)作者简介:张雨薇(1997-),女,硕士研究生,E-mail:501558951@qq.com通讯作者:韩德明(1979-),男,博士,副教授,E-mail:handeming2009@yahoo.com狂犬病病毒aG株假病毒的制备方法张雨薇,韩德明(长春理工大学生命科学技术学院,长春130022)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RVaG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RVaG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RVaG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。关键词:狂犬病病毒;假病毒;aG株;转染中图分类号:R373.9文献标志码:A文章编号:1672-9870(2023)01-0102-06PreparationMethodofRabiesVirusaGStrainPseudovirusZHANGYuwei,HANDeming(SchoolofLifeScienceandTechnology,ChangchunUniversityofScienceandTechnology,Changchun130022)Abstract:Thepreparationmethodofrabiesvirus(RV)aGstrainpseudoviruswasestablishedbasedonHIVlentiviralpackagingsystem.TherecombinanteukaryoticexpressionplasmidpCMV-aG-GencodingRVaGstrainglycoprotein(GP)wasconstructedandtransfectedinto293TcellstoverifytheexpressionofGP.TheGPexpressionplasmid,thepseudoviruspackagingplasmidandthegreenfluorescentprotein(GFP)indicatorplasmidwereco-transfectedinto293Tcellsforpseudoviruspackaging;andtheRVpseudoviruscarryingtheGFPreportergeneandaGstrainGPwasprepared.TheresultsshowedthatarecombinanteukaryoticexpressionplasmidencodingRVaGstrainGPwassuccessfullycon-structedinthisstudy,whichcouldbehighlyexpressedin293Tcells;acell-infectiousRVaGstrainpseudoviruswassuc-cessfullyprepared.ThisstudycontributestosaferandmoreaccuratemonitoringofneutralizingantibodylevelsagainsttheimmobilizedaGstrainofRVusedintheproductionofhumanrabiesvaccinesinChina.Keywords:rabiesvirus;pseudovirus;aGstrain;tran...