【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。更多还原

【Abstract】 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.更多还原