CAJ | 학술논문

目的：构建人钙整合素结合蛋白1(calcium- and integrin-binding protein 1, CIB1)基因 RNA 干扰慢病毒载体,获得稳定下调 CIB1表达的慢病毒。方法根据 CIB1的序列,设计 siRNA 序列,克隆入慢病毒核心质粒 pGV112中,并与辅助质粒一起转染293T 细胞进行病毒包装,用慢病毒感染293T 细胞并测定病毒滴度。以 CIB1干扰慢病毒感染人胶质瘤细胞 SHG44后,应用 Western blotting 方法检测 CIB1的表达。结果成功构建 CIB1干扰慢病毒载体,包装后获得慢病毒颗粒,病毒滴度检测可达6×108 pfu/ ml,感染细胞 CIB1蛋白表达水平显著下调(P <0.01)。结论成功构建人CIB1 基因RNA 干扰慢病毒,为后续CIB1 生物学功能研究奠定基础。
목적：구건인개정합소결합단백1(calcium- and integrin-binding protein 1, CIB1)기인 RNA 간우만병독재체,획득은정하조 CIB1표체적만병독。방법근거 CIB1적서렬,설계 siRNA 서렬,극륭입만병독핵심질립 pGV112중,병여보조질립일기전염293T 세포진행병독포장,용만병독감염293T 세포병측정병독적도。이 CIB1간우만병독감염인효질류세포 SHG44후,응용 Western blotting 방법검측 CIB1적표체。결과성공구건 CIB1간우만병독재체,포장후획득만병독과립,병독적도검측가체6×108 pfu/ ml,감염세포 CIB1단백표체수평현저하조(P <0.01)。결론성공구건인CIB1 기인RNA 간우만병독,위후속CIB1 생물학공능연구전정기출。
Objective To construct a method of lentiviral vector for RNA interference (RNAi) of human calci-um- and integrin-binding protein 1 (CIB1) gene to obtain lentiviral stably down-regulated CIB1 expression. Methods The sequence of siRNA for CIB1 interference was cloned into the pGV112 vector, and viruses packaging with assistant plasmids in 293T cells was performed, and then 293T cells were infected with the lentivirus, the virus titer was detected. The CIB1 expression was detected by Western blotting after human glioma cells SHG44 were infected by CIB1 interfered in lentivirus. Results The lentiviral particle was packaged with a virus titer reaching 6 × 108 pfu/ ml after interference vector CIB1 was successfully constructed, and the level of CIB1 protein expression was significantly down-regulated (P <0. 01). Conclusion The lentiviral vector for RNA interference of CIB1 has been successfully constructed, and it pro-vides a foundation for further research of CIB1 biological function.