内部核糖体进入位点控制人癌胚抗原与IL-2的共表达  

Control of coexpression plasmid of carcinoembryonic antigen and IL-2 gene by internal ribosome entrying site

作　　者：何天霖[1] 傅传刚[1] 王中川[1] 曹贵松[1] 王庆敏[2] 吴丹[2] 戴建新[2] 孙树汉[2] 

机构地区：[1]第二军医大学长海医院普外科 [2]第二军医大学医学遗传教研室,上海200433

出　　处：《中国普通外科杂志》2002年第11期668-670,共3页China Journal of General Surgery

基　　金：国家自然科学基金资助 (30 0 70 744)

摘　　要：目的　构建能在真核细胞内高效表达的癌胚抗原 (CEA ) /IL 2共表达核酸疫苗 ,为进一步研究CEA核苷酸疫苗、佐剂及它们的特异性抗肿瘤作用奠定基础。方法　通过RT PCR及基因重组技术获得CEA的真核表达质粒 pcDNA 3 CEA ,并利用酶切、连接等手段与IL 2基因重组通过内部核糖体进入位点连接的含有两个表达单元的真核共表达质粒 pIRES CEA /IL 2。 结果　全自动电化学法定量测定CEA表达量为 (2 3 .73± 0 .2 6)ng/ml ,ELISA试剂盒测定IL 2表达量为 (2 0 .17± 0 .13 )ng/ml。 结论　CEA /IL 2共表达载体在真核细胞中能高效表达CEA和IL 2 ,并在表达量上无明显差异。ObjectiveTo construct the eukaryotic coexpression plasmid CEA/IL-2, and to lay the foundation for further studying CEA nucleiotide vaccine , adjuvant and their effects of special antitumor immunity.MethodsThe eukaryotic expression plasmid (pcDNA3-CEA)containing the gene coding for CEA was obtained by RT-PCR and gene recombination techniques.Using enzymolysis,ligation and other techniques,an eukaryotic coexpression plasmid (pIRES-CEA/IL-2)containing two expression unites of CEA and IL-2 gene connected with internal ribosome site was constructed.ResultsThe coexpression plasmids were transformed into COS7 cells and expression of two proteins were demonstrated by ELISA, and flow cytometer and elecsy.CEA and IL-2 were (23.73±0.26)ng/ml,and(20.17±0.13)ng/ml respectively.ConclusionsThe eukaryotic expression plasmids pIRES-CEA/IL-2 could be successfully constructed and transformed into COS7 cells.Expression of two proteins were demonstrated with no difference on expression.

关 键 词：癌胚抗原 遗传学 白细胞介素-2 核糖体 基因表达 RT-PCR 基因重组 

分 类 号：R730.3[医药卫生—肿瘤]