一种可药物筛选及体内检测的新型慢病毒基因治疗载体的构建  

Construction of a Novel Gene Therapy Lentiviral Vector for Drug Resistant Selection and Detection in vivo

作　　者：许辰煜[1] 顾颖[1] 侯汪衡[1] 程通[1] 张涛[1] 阙玉琼[1] 高双全[1] 张军[1] 韩家淮[2] 夏宁邵[1] 

机构地区：[1]厦门大学国家传染病诊断试剂与疫苗工程技术研究中心,厦门361005 [2]厦门大学生命科学学院,厦门361005

出　　处：《生物工程学报》2008年第2期256-261,共6页Chinese Journal of Biotechnology

基　　金：福建省科技重大专项资助(No．2004YZ01);~~高等学校科技创新工程重大项目培育资金项目资助(No．705031)~~

摘　　要：为实现基因治疗过程中的有效药物筛选及体内检测,首次利用核糖体内部进入位点(IRES)构建了同时携带O6-烷基鸟嘌呤-DNA烷基转移酶(MGMT)的突变型P140K基因和荧光素酶(Luciferase)基因的慢病毒载体pBobi-MIL。RT-PCR、免疫荧光、药物筛选克隆形成及化学发光检测等实验结果表明感染重组慢病毒L-MIL的细胞能同时表达MGMT及Luciferase。构建成功的新型慢病毒载体为今后的基因治疗奠定了基础,也为慢病毒滴度的确定提供了一种新的可能。Lentiviral vectors were powerful gene delivery tools for gene therapy. We developed a new lentiviral vector pBobi-MIL that constitutively expressed O6-methylguanine-DNAmethyltransferase （MGMT） and Luciferase, linked by the internal ribosomal entry site （IRES）, to realize drug tolerance and real time monitoring in vivo. All results from RT-PCR, drug treating clones forming, immunofluorometric assay and chemiluminescence detection showed that cells infected by recombinant lentivirus L-MIL simultane- ously expressed these two genes. This lays the foundation for the further research in gene therapy and can also help identify lentivirus titer.

关 键 词：基因治疗 慢病毒载体 体内检测 MGMT LUCIFERASE 

分 类 号：Q78[生物学—分子生物学]