FRAT1基因真核表达质粒的构建与鉴定

Construction and Identification of Eukaryotic Expression Vector for FRAT1 Gene

目的构建FRAT1siRNA真核干涉表达载体pSINsi-FRAT1,并测序鉴定证实克隆正确。方法设计以FRAT1基因为靶标的RNAi序列和阴性对照片段序列,并克隆入载体pSINsi-hU6,用基因测序进行重组克隆验证。结果 PCR检测证实siRNA插入pSINsi-hU6质粒,测序分析证实插入序列正确。结论成功构建FRAT1基因的RNAi载体,为研究FRAT1基因对结肠癌细胞增殖凋亡侵袭的影响提供了稳定转染的RNA干扰质粒。

Objective To construct a eukaryotic expression vector for FRAT1 gene,Sequencing proved the correct sequence of this vector. Methods FRAT1 siRNA sequence and the negative control fragmentwas designed,synthesized and cloned into the expression vector pSINsi-hU6. The recombinants were identified by DNA sequencing,Results PCR test and DNA sequence analysis showed that the recombinant plasmid was constructed correctly. Conclusion A vector carrying a siRNA targeting FRAT1 has been constructed successfully,and it provided stable transfection RNA interfer- ence plasmid for the research of FRAT1 participation in coloncells.