结缔组织生长因子小干扰RNA的构建及其干扰效果检测

目的：构建结缔组织生长因子（CTGF）基因的小干扰RNA（siRNA），并检测其对CTGF基因表达的干扰效果。方法：根据CTGF基因序列，设计2条合理的CTGF-siRNA，并将其克隆到siRNA载体pSliencer2．1-U6neo中，转化大肠杆菌DH5a，挑取阳性菌落进行酶切和测序鉴定；将构建成功的CTGF-siRNA重组质粒与带nJAG标签的CTGF表达载体共同转染人胚肾293T细胞，Western印迹检测siRNA的干扰效果。结果：构建了2个CTGFpsiRNA重组质粒，2条siRNA都有干扰作用，其中一条的干扰效果可达75％以上。结论：构建的CTGF-siRNA可为进一步研究CTGF的功能提供参考。

Construction of Small Interfering RNA of Connective Tissue Growth Factor and its Interference Effect

Objective： To construct small interfering RNA（siRNA） of connective tissue growth factor（CTGF） expression vector, and investigate its interference effect. Methods： Two CTGF-siRNA were designed and inserted into the pSliencer 2.1-U6 neo vector. After confirmation of the constructs by restriction and DNA sequencing, human embryo kidney 293T cells were co-transfected with the recombination plasmids and FLAG-CTGF expression vector. The interference effect of CTGF-siRNA was identified by Western blot. Results： Two CTGF-siRNA expression vectors were obtained and could effectively inhibit the expression of CTGF protein, one of which could inhibit the expression over 75%. Conclution： The CTGF-siRNA expression vectors constructed can provide tools for further study on function of CTGF.