慢病毒载体pITA-HBP1的制备及感染效率的测定

Construction and Evaluation of Lentiviral Vector pITA-HBP1

慢病毒载体是目前基因治疗中研究较多的载体,与通常使用的逆转录病毒载体和腺病毒载体比较,它有感染非分裂期细胞及容纳大片段外源性目的基因等优点.本文应用基因重组的方法,构建了1种新型的慢病毒载体,并将转录因子HBP1基因插入到这一载体上,成为pITA-HBP1.检测其在人的肿瘤细胞和正常细胞中的转染效率,发现转染效率明显增加,Western印迹证实外源基因得到高效表达.这一结果,对今后实验室提高基因的转染效率、表达水平,以及研究基因的表达调控,都提供了重要的技术方案.

Lentiviral vectors have been widely used in gene therapy and in laboratory biological study for their ability to deliver larger gene fragments into cells in the non-dividing phase.In this study,we constructed a lentiviral vector pITA-HBP1 expressing the HBP1 transcription factor and evaluated its infection efficiency of the packaged lentivirus by Western blot.We found that both human tumor cells and normal cells were infected efficiently as determined by the expression of HBP1.Our results showed that HBP1 could be transduced exogenously into normal or cancer cells with our lentiviral system.