靶向survivin基因的siRNA真核表达载体的构建及鉴定  

Construction and identification of eukaryotic expression vector of target survivin

作　　者：李红[1] 李明红[1] 缪书卉[1] 杨桦[1] 镡旭民[1] 

机构地区：[1]第三军医大学新桥医院耳鼻咽喉头颈外科,400037

出　　处：《重庆医学》2008年第11期1184-1185,1188,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(No30672299)

摘　　要：目的构建靶向survivin基因的siRNA的表达载体,研究survivin在喉癌中的作用机制。方法根据基因库上sur-vivin cDNA序列,设计合成靶基因序列,将其寡核苷酸退火后插入到Pgenesil-1表达载体中,对该重组表达载体进行酶切鉴定及DNA测序。结果酶切鉴定、DNA测序证实表达质粒构建成功,无碱基突变。结论成功构建了靶向survivin基因表达的siR-NA干扰质粒载体Pgenesil/survivin,为进一步运用RNA干扰技术进行survivin基因功能研究奠定了基础。Objective To construction the survivin-siRNA expression vector and to research the function of survivin in laryngeal squamous cell carcinoma. Methods According to survivin cDNA sequence in genebank,the target sequence was designed and synthesized. After annealing, oligonucleotides were inserted into pGenesil-1 vector, the recombined plasmid was identified by restriction endonuclease and DNA sequencing. Results By restriction endonuclease, eukaryotic expression plasmid of survivin proved to be successfully constructed. Sequence analysis of inserted fragment revealed the same sequence as synthesized oligonucleotides. Conclusion The survivin-siRNA expression vector has been successfully constructed. It is foundation for studying the function of survivin by small interfere RNA technique.

关 键 词：SURVIVIN基因 小干扰RNA 质粒 喉癌 

分 类 号：R739.6[医药卫生—肿瘤]