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Construction of mammalian co-expression plasmid pIRES-CEA- IL-2 and its expression in vitro
Author(s): 
Pages: 193-196
Year: Issue:  3
Journal: CHINA ONCOLOGY

Keyword:  DNA疫苗癌胚抗原共表达质粒白细胞介素2;
Abstract: 目的:构建癌胚抗原(CEA) 与白细胞介素2(IL-2)的真核双表达质粒,并检测其在体外的表达.方法:利用分子生物学技术,将CEA基因片段和IL-2基因片段连接于真核双表达质粒pIRES中,测定序列后,用脂质体包裹转染细胞,采用ELISA双抗体夹心法检测CEA和IL-2的表达.结果:核酸序列测定证实本实验所构建的质粒正确,质粒上所接入的CEA和IL-2的碱基序列与标准序列的同源性分别为99.8%和99.9%.该双表达质粒在体外转染细胞内后可表达CEA和IL-2分子.结论:实验所构建的pIRES-CEA- IL-2双表达质粒能在体外同时表达CEA和IL-2分子,说明本质粒具有在细胞水平同时表达两种生物大分子的活性,这一结果为其以后在整体动物水平的实验研究奠定了基础.
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