中文摘要：

目的构建内皮抑素（endostatin,ES）过表达慢病毒载体,并进行鉴定。方法利用载体PUC57-ES构建GV365-ES慢病毒载体,转染293T细胞包装慢病毒,荧光显微镜下观察绿色荧光;Western blot法鉴定表达的目的蛋白;HIV-1 ELISA法检测病毒滴度。结果经PCR及测序鉴定证明慢病毒载体GV365-ES构建正确;转染细胞中荧光显微镜下可见绿色荧光,表达的目的蛋白相对分子质量约23 000;病毒滴度为2.1×10^8 TU/ml。结论已成功构建ES过表达慢病毒载体,为后期该病毒载体治疗实体瘤的观察及其作用机理的研究奠定了基础。

英文摘要：

Objective To construct and identify a lentiviral vector for over-expression of endostatin（ES）. Methods Lentiviral vector GV365-ES was constructed by using plasmid PUC-ES, with which 293 T cells were transfected and observed for green fluorescence by fluorescent microscopy. The expressed target protein was identified by Western blot,and the virus titer was determined by HIV-1 ELISA. Results Both PCR and sequencing proved that the lentiviral vector was constructed correctly. Green fluorescence was observed in the transfected cells under fluorescent microscope. The relative molecular mass of expressed target protein was about 23 000, while the virus titer was 2. 1 × 10^8 TU / ml.Conclusion The lentiviral vector for over-expression of ES was successfully constructed, which laid a foundation of study on mechanism of the lentivirual vector in treatment of solid tumor.