摘要
目的筛选鉴定针对Hl V-1vpr基因的小干扰RNA(siRNA)片段。方法根据siRNA设计要求合成siRNA56、siRNA160和siRNA185寡核苷酸片段,分别转染至含HIV-1vpr质粒的HEK 293T细胞,并进行总RNA提取,采用Real-time PCR和Western blotting分别从核酸和蛋白水平对HIV-1vpr基因表达水平进行验证。结果siRNAs成功转染含HIV-1vpr质粒的HEK 293T细胞,降低了HIV-1vpr基因的表达水平,其中在RNA水平siRNA160组干扰抑制作用最强,抑制率为89%;在蛋白水平siRNA56组干扰抑制作用最强,抑制率为96%。结论 3个基因片段的siRNA均可以下调HIV-1vpr的表达水平,但存在差异性,为探索HIV/AIDS基因治疗的可行性和高效性提供了可靠的实验依据。
Objective To screen and identify fragments of siRNA on HIV - 1vpr gene. Methods This study designed and synthesized siRNA56,siRNA160 and siRNA185 oligonucleotide fragments according to siRNA design requirements, transfected them into HEK293T cells containing plasmids of HIV - 1vpr genes,extracted total RNA and verified HIV - 1vpr from levels of nucleic acid and protein by Real - time PCR and Western blotting. Results The siRNA successfully transfected HEK 293T cells containing HIV - 1vpr plasmids and reduced the expression of HIV - 1vpr genes. The inhibition rate of siRNA160 group was the highest in RNA level（89% ）and that of siRNA56 group was the highest in protein level（96% ）. Conclusion The siRNAs of three gene fragments,which can decrease the expression HIV - 1vpr,provide a reliable and efficient experimental basis for exploration of HIV/ AIDs gene treatment.
出处
《中国全科医学》
CAS
CSCD
北大核心
2015年第14期1671-1674,共4页
Chinese General Practice
基金
湖南省科学技术厅科技计划项目(2014SK3097)
