人GOR基因重组体的构建及其在C_(os)—7细胞内的表达  

Constructing the recombinant of the humanbeings' GOR gene and its expression within Cos-7 cell line

作　　者：何剑琴[1] 万群[1] 陈智[1] 章明太[1] 刘克洲[1] 徐峰[1] 

机构地区：[1]浙江大学医学院附属第一医院,杭州310003

出　　处：《中华肝脏病杂志》1999年第S1期36-38,共3页Chinese Journal of Hepatology

摘　　要：目的构建能正确表达人GOR蛋白的真核重组体，制备人源性GOR蛋白。方法将人GOR基因片段亚克隆到真核表达载体PCDNA31（－）／Myc－HisA，B，C系统，PCR法和EC。RⅠ酶切筛选重组体，KpnⅠ酶切鉴定重组体方向，正向重组体用磷酸钙共沉淀法分别转染Cos－7细胞。用PCR法和ELISA法分析重组体是否转入Cos—7细胞并能正确编码人GOR蛋白。最后制备人GOR蛋白。结果所构建的人GOR基因重组体A、B、C分别转染Cos—7细胞后，都能在细胞内转录；但只有重组体A能正确表达人GOR蛋白。结论　本研究所构建的人GOR基因真核重组体能正确表达人GOR蛋白。Objective To construct eukariotic recombinants able to express human beings' GOR protein and to abtain the protein. Methods The fragments of human being's GOR gene were inserted to the vector system of pcDNA3.1(-)/Myc-His A, B and C, and then positive recombinants were selected by the technique of PCR and EcoR I digestion. The recombinants with sense sequense were determined by Kpn I digestion. After three kinds of recombinant transfecting Cos-7 cells with the calcium phosphate precipitation method respectively, the technique of PCR and ELISA were used to confirm whether the recombinants could be transcripted and expressed or not within cells. Plenty of Cos-7 cells cultured and transfected with recombinant-A, the protein of human beings' GOR was got. Results All of the three kinds of recombinant A, B, C could be transcripted within Cos-7 cells, but only recombinant A could express human beings'GOR protein. Conclusion Eukariotic recombinants constructed could exactly express human beings' GOR protein. [

关 键 词：丙型肝炎 自身免疫 抗-GOR抗体 

分 类 号：R394[医药卫生—医学遗传学]