【作者】 王代友； 于大海； 邝晓聪； 张斌；

【Author】 Wang Da-iyou, Yu Da-hai, Kuang Xiao-cong, Zhang Bin The department of oral maxillofacial surgery, affliliated stomatological hospital, Guang Xi Medical university

【机构】 广西医科大学附属口腔医院口腔颌面外科；

【摘要】 <正>目的:克隆人癌胚抗原基因cDNA,构建癌胚抗原DNA疫苗。方法:(1)基因疫苗的构建:采用RT-PCR 技术,从人结肠癌组细胞中克隆出癌胚抗原(carcinoembryonic antigen,CEA)基因片段,通过DNA重组技术将该基因片段重组于pcDNA,真核表达载体上,构建成pcDNA3／CEA基因疫苗。(2)基因疫苗的鉴定:分别用PCR扩增、酶切电泳分析及DNA测序的方法对pcDNA3／CEA基因疫苗进行鉴定。结果:经 PCR扩增可见2．1kb目的基因片段;酶切电泳分析可见2．1kb目的基因片段和5．4kbpcDNA3质粒片段;更多还原

【Abstract】 Objective: The aim of this study is to clone CEA gene and to compose the expressive plamsid pcDNA3/CEA Methods: CEA gene was cloned by RT-PCR method from human colon cancer and then was composed into the expressive plamsid pcDNA3/CEA. The gene in the expressive plamsid pcDNA3/CEA is identified by PCR amplifcation, enzyme digestion and DNA sequencing. Results: A bar of 2.1kb occured in the electrophoretic path by PCR amplification, two bars of 2.1kb and 5.4kb were found in the same electrophoretic path by enzyme digestion, and the sequence of the cloned cDNA was completely consistent with the sequence of CEA cDNA by DNA sequencing. Conclusion: In this study we successfully cloned CEA gene and composed its pcDNA3/CEA gene vaccination, which provided the foundation for our further study of antitumor by pcDNA3/CEA gene vaccination.更多还原