MMP-24基因siRNA真核表达载体的构建及鉴定

The Establishment and Identification of siRNA Eukaryotic Expression Vectors of Gene MMP-24

目的:构建基质金属蛋白酶-24(matrix metalloproteinase-24,MMP-24)小干扰RNA(small interfering RNA,siRNA)表达质粒,以研究其对人卵巢浆液性囊腺癌细胞MMP-24基因表达的沉默效果,为探讨肿瘤的基因治疗奠定基础。方法:根据Gen-bank数据库提供的MMP-24基因核苷酸序列,选择设计能转录小发卡RNA(small hairpin RNA,shRNA)结构的DNA序列,克隆到空载体pIU6-B中,构建重组质粒,并进行酶切和PCR鉴定以及测序分析。结果:酶切和PCR鉴定以及测序证实重组质粒构建成功。结论:利用RNA干扰技术可成功构建siRNA表达载体,为进一步探索卵巢癌基因治疗奠定了基础。

Objective： To establish the recombinant plasmid of small interfering RNA （siRNA） against matrix metalloproteinase-24（MMP-24）, observe dumbness effect of the MMP-24 gene expression in human ovarian serous cystadenocarcinoma cells, and explore the new method of gene therapy. Methods： Genmoe se- quences of MMP-24 gene was retrieved from Genbank and cDNA was designed coding expression of shRNA （small hairpin RNA） for MMP-24 gene. The cDNA was synthesized and inserted into plasmid pIU6-B to generate siRNA eukaryotic expression vectors, and the recombinant sequences were identified. Results： The result of recombinant sequence was the same as aim sequence. The recombinant vectors were established successfully. Conclusion： siRNA recombinant can be established successfully by RNA interference（RNAi） technique, which will facilitate further studies of MMP-24 function and its application in the treatment of overian carcinoma.