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.