真核双基因表达载体pIRES-IL-24-ES的构建与鉴定  

Construction and identification of the eukaryotic co-expression plasmid pIRES-IL-24-ES

作　　者：韩建军[1] 胡三元[1] 智绪亭[1] 梁晓红[2] 薛德文[3] 

机构地区：[1]山东大学齐鲁医院普通外科,济南250012 [2]山东大学医学院免疫学实验室,济南250012 [3]山东省肿瘤医院肿瘤介入治疗中心,济南250117

出　　处：《山东大学学报（医学版）》2010年第3期78-82,共5页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金项目(Y2007C150)

摘　　要：目的构建白介素24(IL-24)与内皮抑素(ES)的真核双基因表达载体,检测它们在体外的表达。方法从外周血和人胎肝组织中经RT-PCR分别扩增IL-24与EScDNA序列,并将其定向克隆至真核双基因表达质粒pIRES中,重组质粒pIRES-IL-24-ES,经酶切及测序鉴定后,转染NIH3T3成纤维细胞。经RT-PCR及ELISA法检测IL-24和ES在NIH3T3中的表达。结果重组真核双基因表达载体pIRES-IL-24-ES构建成功,IL-24与ES可在NIH3T3细胞中有效表达。结论真核双基因表达载体pIRES-IL-24-ES成功构建为研究肿瘤的基因治疗提供了一定线索。Objective To construct the eukaryotic co-expression plasmid encoding interleukin-24（IL-24） and endostatin（ES）,and to detect expression of the plasmid in vitro.Methods The cDNA encoding Il-24 and ES were obtained by RT-PCR amplifications from peripheral blood and normal fetus liver respectively and cloned into an eukaryotic expression plasmid internal ribosome entry site（pIRES）.The co-nstructed plasmid pIRES-IL-24-ES was confirmed by restriction enzymolysis and DNA sequencing.Fibroblast NIH3T3 cells were transformed by plasmid pIRES-IL-24-ES.RT-PCR and ELISA were used to identify the co-expression of IL-24 and ES in NIH3T3 cells.Results The eukaryotic expression plasmid pIRES-IL-24-ES was successfully constructed and the plasmid could efficiently co-express IL-24 and ES in NIH3T3 cells.Conclusion The successful construction of eukaryotic expression plasmid pIRES-IL-24-ES establishes an experimental basis for cancer treatment.

关 键 词：白介素-24 内皮抑素 分泌表达载体 肿瘤 

分 类 号：R392.1[医药卫生—免疫学]