人IL-2基因克隆及其重组腺病毒载体的构建  

Cloning of human Interleukin 2 gene and the construction of recombinant adenovirus vector

作　　者：李安娜[1] 匡志鹏[1] 罗小玲[1] 谢裕安[1] 吴继宁[1] 尚俊英[1] 楼建军[1] 

机构地区：[1]广西医科大学附属肿瘤医院生物医学研究中心,南宁530021

出　　处：《中国医学文摘（肿瘤学）》2008年第1期82-83,共2页Journal of Chinese Medical Abstracts·Oncology

基　　金：广西自然科学基金项目（合同编号：桂科自0640158）

摘　　要：目的克隆人白细胞介素2基因（hIL-2），构建其重组穿梭载体pAdBM5-GFP—hIL-2。方法：应用PHA体外刺激培养的Jurkat细胞以促进hIL-2基因表达．设计特异性引物，应用RT—PCR法扩增hIL-2基因。将测序正确的片段用PmeⅠ和BglⅡ双酶切定向插入到pAdBM5-GFP穿梭载体中，酶切鉴定。结果：RT—PCR方法，成功克隆了hIL-2基因，并构建出pAdBM5-GFP-hIL-2真核表达载体．克隆的hIL-2序列全长528bp，含462个碱基开放读框．结论；构建含人白细胞介素2基因片段的重组腺病毒表达载体（pAdBM5-GFP—hIL-2），为下一步扩增重组腺病毒开展肝癌的免疫与基因治疗研究奠定基础．Objective, To Clone human Interleukin-2（hIL-2） gene from Jurkat tumor cells and construct a recombinant adenovirus vector pAdBM5- GFP-hIL-2. Methods： Specific primers was designed , Human interleukin-2 gene was cloned by by RT-PCR method . the correct sequence hIL-2 gene was directional constructed into pAdBM5 shuttle vector after digestion by PmeI and BgllI, then the recombinant pAdBM5-GFP-hIL-2 was indentificated. Resalts： hIL-2 gene was cloned correctly and recombinant adenovirus shuttle vector pAdBM5-GFP-hIL-2 was constructed successfully. Conclusion, The constructed recombinant adenovirus vector pAdBM5-GFP-hIL-2 will lay down the basis for further amplifying the related recombinant adenovirus and developing gene therapy of primary liver cancer.

关 键 词：人白细胞介素2 真核表达载体构建 pAdBM5-GFP—hIL-2 基因治疗 

分 类 号：R394[医药卫生—医学遗传学] R392.11[医药卫生—基础医学]