目的:以βCalT7(人β1,3-半乳糖基转移酶7)基因为研究对象,运用RNA干扰技术平台,筛选出该基因的有效小干扰siRNA并构建该基因的RNA干扰表达载体,建立其下调表达模型。方法:通过PCR方法筛选针对β3CalT7基因的有效小干扰siRNA,并合成带有荧光标记的RNA oligo进一步验证其有效性,进而构建该基因的RNA干扰表达载体建立其下调表达模型。结果:成功筛选到团β3GalT7基因的有效小干扰siRNA,并进一步成功构建β3CalT7的RNA干扰真核表达载体。结论:成功构建β3CalT7的真核干扰表达载体,继而可以获得β3CalT7表达受抑制的人胃癌细胞SGC7901克隆,为研究该基因在肿瘤发生发展中的作用提供实验模型。
Objective: Based on the RNAi (RNA interfering)technology to construct the knockdown model of β3CalT7 gene. Methods: Using PCR method to screen the most effective small interference RNA of β3CalT7, synthesised fluorescently-labeled RNA oligos to detect its effectiveness and constructed the RNAi-vector of β3CalT7 Results: The knockdown model of β3CalT7 gene was successfully constructed. Conclusion: The constructed RNAi-vector of β3CalT7 lay foundation to functional study of the gene.