HMGN2基因干扰质粒构建及干扰效率测定

Construction and identification of interference plasmid of HMGN2

目的:构建HMGN2基因的干扰质粒,为深入研究HMGN2基因在信号通路中的作用提供有效手段。方法:根据文献选择两个HMGN2基因的干扰位点,合成两个干扰片段定向克隆到psilencer-4.1的干扰载体并测序验证。将干扰质粒转染至A549细胞,并通过检测RT-PCR产物量以获得干扰效率。结果:RT-PCR产物检测结果显示干扰质粒psilencer-4.1-HMGN2-2干扰效率最高,其HMGN2表达量降低了70%左右。结论:成功构建了对HMGN2基因具有显著干扰效率的psilencer-4.1-HMGN2干扰质粒,为进一步研究HMGN2基因的功能打下了基础。

Objective： To construct highly efficient interference plasmid aiming at HMGN2 gene, and provide an effective way for studying the function of HMGN2 gene. Methods：Two interference sites of HMGN2 gene were selected from documents and two interference fragments and one negative control fragment were synthesized, then they were cloned into the psilencer-cmv-4. 1 plasmid. After sequencing, the plasmids were transfected into A549 cells. The interference efficiency of the plasmid on the target gene was detected by semiquantitative RT-PCR. Results： The level of HMGN2 mRNA in A549 ceils contained interference plasmids decreased by 70% compared with negative control plasmid. Conclusion： The effective interference plasmid against HMGN2 gene had been constructed and identified successfully, which may provide a basis for studying the function of HMGN2 gene in signal transduetion pathway.