NS特异性干扰载体pGenesil-1-NS的构建及鉴定  

Construction and Identification of p Genesil-1-NS Interference Vector

作　　者：高鸿霞[1] 王国庆[1] 刘燕[1] 霍中华[2] 李钰[2] 

机构地区：[1]北华大学医学检验学院,吉林吉林132013 [2]北华大学附属医院,吉林吉林132011

出　　处：《北华大学学报（自然科学版）》2015年第6期729-732,共4页Journal of Beihua University（Natural Science）

基　　金：吉林省卫生厅基金项目(2012Z032/2014Z077);吉林市科技创新服务平台项目(2013625027)

摘　　要：目的核干细胞因子(nucleostemin,NS)是维持干细胞和癌细胞增殖所必需的蛋白质,可能成为肿瘤基因治疗的潜在靶点.本文旨在构建靶向NS干扰载体p Genesil-1-NS,为后续实验奠定基础.方法基于已发布的NS mRNA序列(NM_206825),选取5'-AAGCCTA GGAAAGACCCAGG-3'(397-416)作为候选靶序列,按shRNA载体设计原则,设计合成两条互补DNA链,退火后插入载体,并以酶切和测序鉴定重组转化子.结果经酶切及测序鉴定证实合成序列正确插入载体.结论成功构建NS特异性干扰载体p Genesil-1-NS,可用于NS在肿瘤中的功能研究.Objective Nucleostemin,a protein to maintain the proliferations of stem cell and cancer cell, may become a potential target for tumor gene therapy. This study aimed to construct the NS specific interference vector pGenesil-1-NS,and to lay a foundation for further study. Method Based on the published NS-mRNA sequence （NM_206825）,5′-AAGCCTA GGAAAGACCCAGG-3′（397-416） was selected as candidate target sequence. According to the shRNA carrier design principle,shDNA that could be transcripted to shRNA was designed and synthesized,then inserted into pGenesil-1 vector. The recombinant was identified by restriction enzyme digestion analysis followed by sequence analysis. Results Confirmed by restriction analysis and sequencing,shDNA was correctly inserted into the vector. Conclusion NS-target interference vector pGenesil-1-NS was successfully constructed,which would be applied in the study of NS function in tumor generation and progression.

关 键 词：核干细胞因子基因 pGenesil-1载体 发夹状RNA RNA干扰 

分 类 号：Q78[生物学—分子生物学]