人IL-21cDNA的克隆及重组腺病毒表达载体的构建  

CLONING OF IL-21 GENE AND CONSTRUCTION OF RECOMBINANT IL-21 GENE ADENOVINES EXPRESSION VECTOR

作　　者：王芹[1] 李进[1] 王颖嫒[2] 宋力[1] 岳井银[1] 穆传杰[1] 樊飞跃[1] 

机构地区：[1]中国医学科学院放射医学研究所天津市分子核医学重点实验室,天津300192 [2]南方医科大学,广州510515

出　　处：《广西医科大学学报》2009年第4期516-518,共3页Journal of Guangxi Medical University

基　　金：天津市自然科学基金资助项目(No.09JCYBJC09300);中国医学科学院放射医学研究所基金资助项目(No.ST0721)

摘　　要：目的:构建含人IL-21基因的重组腺病毒表达载体,为IL-21基因治疗肿瘤研究奠定基础。方法:从人外周血淋巴细胞提取总RNA,经RT-PCR扩增IL-21基因片段,将其连接到进入载体pENTR1A,然后经LR重组转入到腺病毒表达载体pAd/CMV/V5-DEST中,构建含IL-21基因的腺病毒载体(Ad-IL-21),并进行测序鉴定和PCR鉴定。结果:Ad-IL-21表达载体测序结果与GenBank中IL-21基因序列一致,Ad-IL-21经PCR扩增出IL-21基因片段。结论:成功构建携带人IL-21基因的重组腺病毒表达载体。Objective.To be as the foundation of the tumor therapy with targeting IL-21 gene by constructing of recombinant interleukin 21 gene adenovirus expression vector. Methods. Total RNA was extracted from human peripheral blood lymphocyte. IL-21 gene was amplified by RT-PCR and linked with pENTR1A vector. The constructed vector pENTRIA- IL-21 and adenovirus vector pAd/CMV/VS-DEST were recombined by LR recombinant technique to construct the recombined IL-21 gene adenovirus expression vector（Ad-IL- 21）. Ad-IL-21 was identified by DNA sequencing and PCR method. Result.DNA sequence of IL-21 gene of Ad- IL-21 adenovirus vector was in accordance with that of IL-21 gene in GenBank database. IL-21 gene fragment of Ad- IL-21 vector was amplified by PCR. Conclusion.The recombinant Ad-IL-21 expression vector could express IL-21 gene and was constructed successfully.

关 键 词：IL-21基因 基因克隆 腺病毒表达载体 LR重组反应 

分 类 号：R730.5[医药卫生—肿瘤]