中文摘要：

目的构建携带小鼠血管生成素-1（Ang-1）基因的慢病毒载体并进行慢病毒包装。方法利用Gateway技术构建携带DsRed荧光蛋白的Ang-1慢病毒载体，经PCR及基因测序鉴定后，与辅助质粒pLV／helper-SL3、pLV／helper-SL4及pLV／helper-SL5混合采用脂质体法制备DNA-Lipofectamine2000复合物，并共同转染293FT细胞进行幔病毒包装，产生相应慢病毒颗粒，通过荧光表达情况来测定病毒滴度和感染效率。结果通过PCR、基因测序证实，Ang-1慢病毒表达载体PLV．Ex3d．P／puro-CMV〉Ang-1〉IRES／DsRed-Express2构建成功。经转染293FT细胞包装出具高效感染力的慢病毒，经测定病毒滴度为5×10^8 TU／ml。结论成功构建Ang-1慢病毒表达载体，并包装出具高效感染力的慢病毒颗粒，为进一步研究Ang-1基因功能及基因治疗提供实验基础。

英文摘要：

Objective To construct a lentiviral vector carrying angiopoietin-1 （Ang-1） and DsRed gene, and to package a virus particles. Methods The Ang-1 lentiviral vector with DsRed （PLV.Ex3d.P/puro-CMV〉Ang-l〉IRES/DsRed-Express2） was constructed by Gateway technology, and identified by PCR and gene sequencing. The lentiviral vector was mixed with helper vector pLV/helper-SL3, pLV/helper-SL4 and pLV/helper-SL5 by Lipofectamine 2000 to prepare DNA-LipofeetamineTM 2000 complexes. The complexes were then added to transfect 293FT cells and package virus. The virus titers and infection ef- ficiency were determined by fluorescence expression. Results Ang-1 lentiviral vector PLV.Ex3d.P/puro-CMV〉Ang-1〉IRES/ DsRed-Express2 was constructed successfully as identified by PCR and gene sequencing. Lentivirus with high-efficiency infection was produced by transfection to 293FT cells and the virus titer was 5×10^8 TU/ml. Conclusions The recombinant len- tiviral vector for Ang-1 was successfully constructed by Gateway technology and the lentivirus with high-efficiency infectionpackaging can be used for further experiment orang-1 gene.