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

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.