摘要
目的:构建核因子-κB(nuclear factor-κB,NF-κB)p65亚基特异的核糖核酸干扰腺病毒表达载体,验证其对p65亚基的沉默效应。方法:设计针对大鼠NF-κB p65亚基的不同干扰序列,通过质粒将其导入腺病毒,感染大鼠心肌细胞(H9C2)。将感染后的心肌细胞通过RT-PCR方法和Western Blot方法检测各组NF-κB p65的基因水平表达和蛋白质水平表达。结果:成功构建了NF-κB p65亚基特异的RNAi腺病毒表达载体,获得高滴度的腺病毒液;RNAi腺病毒感染H9C2细胞后可以高效抑制NF-κB p65蛋白的表达。结论:RNAi技术能有效的沉默大鼠心肌细胞NF-κB p65的表达。
Objective：Construction of NF-κB p65 subunit-specific RNA interference adenovirus expression vector,Verify its effect on the p65 subunit of silence.Methods：Design different interference sequences for rat NF-κB p65 subunit,import it to adenovirus by plasmid,and then transfected the adenovirus into rat cardiomyocytes.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and TRPCR in rat cardiomyocytes.Results：The NF-κB p65 subunit-specific RNAi adenovirus expression vectors was successfully constructed and the high titer adenovirus venom was obtained; The H9C2 cells infected by RNAi adenovirus can efficiently inhibit the NF-κB p65 protein expression.Conclusion：RNAi technology can effectively silence NF-κB p65 expression in rat cardiomyocytes.
出处
《心肺血管病杂志》
CAS
2014年第2期267-271,共5页
Journal of Cardiovascular and Pulmonary Diseases
基金
北京市自然科学基金(7112043)
