目的 构建小鼠RPS20 miRNA干扰载体，进行载体质粒的提取及鉴定，以进行细胞转染实验。方法 针对小鼠RPS20基因miRNA序列，设计合成阴性对照及4对目标基因oligo，采用载体构建试剂盒进行重组克隆，经过退火、连接及转化后，进行测序及质粒提取鉴定。结果 测序结果提示序列构建完全正确，电泳结果提示质粒大小正确，纯度较高。结论 成功构建了针对于小鼠RPS20基因的miRNA干扰载体，测序及电泳结果提示，所构建序列可用于后续转染实验。

Objective RPS20 miRNA interference vector was constructed，and vector plasmid was extracted and identified for supplying evidence for cellular transfection experiment. Methods According to mRNA sequence of mouse RPS20 gene，we designed and synthesized the negative reference and four pairs of target genes（oligo）. Vector construction kit was used for obtaining recombinant clones，and then the sequencing and identification of recombinant clones and plasmid extract were performed after annealing，connection，and transformation. Results The sequencing results suggested that the constructed sequence of the plasmid was entirely correct，and the results of electrophoresis suggested the digested plasmid had correct size and higher purity. Conclusion Mice RPS20 miRNA interference vector has been successfully constructed，and the results of sequencing and electrophoresis suggested that the constructed sequences can be used to subsequent transfection experiments.

R349.6；R285.5

国家自然科学基金面上项目（90809001）；中央财政支持地方高校专项资金项目“难治性脾胃病防治协同创新平台（财教[2013]338号）”。