2型腺伴随病毒基因治疗载体的构建

目的　构建一种适用于中枢神经系统疾病的基因治疗载体。方法　以野生型 2型腺伴随病毒 (AAV 2 )质粒pssv9int 及逆转录病毒载体质粒plxsn为基本元件 ,利用重组DNA技术 ,构建了一种通用型AAV载体质粒pssHG Neo。结果　该质粒除含有必备的原核细胞复制子及Ampr 筛选基因外 ,还含有真核细胞筛选基因———Neor 及供插入目的基因的多克隆位点 (MCS) ,在MCS的上游为逆转录病毒的长末端重复序列 (5’LTR) ,具有启动子的功能 ,启动定向插入到MCS的外源目的基因。质粒中的AAV反向末端重复序列 (ITR)具有定点整合于人基因组 1 9号染色体S1位点的特点。结论　AAV载体是具有广阔前景的基因治疗载体

Gene therapy vector based on adeno-associated virus type-2

Objective To construct an adeno associated virus vector(AAV) plasmid used for gene therapy of central nervous system diseases. Methods By using recombinant DNA techniques, a universal adeno associated virus vector pssHG Neo was constructed,which was derived from a wild type adeno associated virus type 2 plasmid pssv9int and a retrovirus plasmid plxsn.Results This vector plasmid contained procaryotic cells propagating and selecting genes: PUC origin and ampicillin resistance (Amp r) gene.It also contained a multiple cloning sites(MCS) and an eucaryotic cells' selecing gene: neomycin resitance (Neo r) gene. 5'LTR (long terminal repeat sequence) of retrovirus,which was located at the upstream of MCS,functioned as a promoter to control the foreign gene inserted in MCS. Since this plasmid flanked with AAV ITRs (inverted teminal repeat sequence),the foreign gene will be stably integrated into specific locus of human chromosome 19 (AAVS1).Conclusion AAV vectors will be promising gene therapy vectors.