CD147 siRNA表达载体的构建与鉴定  

Construction and Identification of CD147 siRNA Expression Vector

作　　者：陈翔[1] 颜克香[1] 谢红付[1] 李吉[1] 

机构地区：[1]中南大学湘雅医院皮肤科,长沙410008

出　　处：《中华皮肤科杂志》2005年第6期368-370,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金(30200248);教育部留学回国人员科研启动基金;教育部高等院校青年教师基金(2003-189);霍英东高校青年教师基金(91040);湖南省自然科学基金(02jjy4018)

摘　　要：目的构建CD147siRNA表达载体,分析CD147在皮肤肿瘤浸润和转移中的作用机制。方法根据基因库上的CD147cDNA序列,设计并合成两端含有酶切位点的64个碱基的寡核苷酸链。寡核苷酸链退火后用T4DNA连接酶连接到线性化的pSUPER质粒中,并对重组质粒(命名为pSUPER/CD147siRNA)进行酶切、PCR及测序鉴定。结果PCR扩增片段与预期结果相符,双酶切证实CD147siRNA表达载体克隆构建成功,插入片段测序结果与合成的siRNA结果一致。结论成功构建CD147siRNA表达载体,CD147可为治疗肿瘤开辟新途径。Objective To construct CD147 siRNA expression vector in o rder to analyze the role of CD147 in the invasion and metastasis of tumors deriv ed from skin.Methods According to CD147 cDNA sequence in the Genebank,a pair of 64-nt oligonucleotides,each containing the sites of restriction endonucleas e at both ends,were designed and synthesized.Oligonucleotides were annealed an d ligated with linearized pSUPER by T4DNA ligase.The recombinants (named pSUPER /CD147 siRNA ) were finally sequenced and identified by PCR and restriction end onuclease digestion.Results CD147 siRNA expression vector was successfully co nstructed and identified by double endonuclease digestion.Sequence analysis of inserted fragment revealed the same sequence as synthesized siRNA oligonucleotid es.Conclusions CD147 siRNA expression vector has been successfully constructed,which paves the way for studying the role of CD147 in the invasion and metasta sis of tumor cells derived from skin as well as in tumor therapy.

关 键 词：CD147 siRNA 表达载体 鉴定 T4DNA连接酶 PCR扩增片段 CDNA序列 寡核苷酸 浸润和转移 作用机制 皮肤肿瘤 重组质粒 插入片段 基因库 酶切 R质粒 线性化 合成 测序 

分 类 号：R739.5[医药卫生—肿瘤]