摘要
目的构建靶向survivin siRNA真核表达质粒载体,通过RNA干扰技术阻断survivin基因的表达。方法针对目的基因survivin序列设计双链DNA(dsDNA),采用DNA重组技术与载体连接,转化感受态细胞经诱导表达筛选阳性克隆子。结果重组质粒经酶切、测序鉴定,证实插入载体目的基因片段大小与插入方向正确,且转染至肿瘤细胞,可见其发出绿色荧光,转染率为70%-80%。结论靶向survivin-siRNA真核表达载体pGU6/GFP/Neo/survivin-siRNA可用于转染肿瘤细胞、阻断survivin基因表达并诱导细胞凋亡的RNA i实验研究,为肿瘤的基因治疗奠定一定的基础。
Objective A eukaryotic expression vector targeting on survivin siRNA is constructed to block up the expression of survivin gene through RNA interference techniques. Methods The double chain DNA （dsDNA） was designed upon the objective gene survivin sequence. The dsDNA was connected with the vector by the DNA recombination techniques. And the sensitive cell was induced to expression screening positive clone cell. Results After the enzyme cutting and sequence identification, it was verified that the size and direction of objective gene segment inserting vector. And transfected into tumor cells, it emited green fluorescence,the transfection rate was 70%-80%. Conclusions Eukaryotic expression vector pGU6 / GFP/Neo/survivin-siRNA targeting on survivin siRNA can be used in the RNAi experimental research of transfecting tumor cells, interrupting the expression of survivin gene and inducting apoptosis. So it can set foundation for the gene therapy of tumor.
出处
《遵义医学院学报》
2008年第6期565-567,570,共4页
Journal of Zunyi Medical University
基金
贵州省科技厅基金(黔科合J字[2007]2128号)
