表达乙肝病毒表面抗原人参细胞系统的建立  

Construction of Expression System of HBsAg in Ginseng Cells

作　　者：刘丹[1] 盛军[1] 刘晓宇[1] 于海鹏[2] 王志武[1] 李娟[1] 姚为民[1] 杨贵贞[2] 

机构地区：[1]长春生物制品研究所,长春130062 [2]吉林大学,长春130023

出　　处：《中国生物制品学杂志》2005年第6期454-456,共3页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划项目(No.20030405)资助

摘　　要：目的建立乙肝病毒表面抗原(HBsAg)的人参细胞表达系统。方法构建含有HBsAg基因的植物细胞表达载体质粒pBIBSb。采用农杆菌感染人参细胞的方法,经农杆菌培养、农杆菌与受体细胞共培养、受体细胞的脱菌培养以及转化体细胞的选择培养后,筛选在G418抗生素培养基上正常生长的细胞株。应用ELISA方法检测抗性细胞株的HBsAg表达;提取基因组DNA进行PCR扩增反应。结果质粒pBIBSb的酶切结果正确。经G418抗生素筛选得到8株正常生长的细胞株,其中5株能够表达HBsAg,提取基因组DNA进行PCR扩增反应,得到大小约700 bp的扩增片段。结论获得了整合HBsAg基因,并能够稳定表达HBsAg的人参细胞表达系统。Objective To construct an expression system of HBsAg in ginseng ceils. Methods Construct a recombinant plasmid pBIBSb carrying HBsAg gene and transform to Agrobacterium tumefaciens strain LBA4404. Co-cultivate the recombinant strain with ginseng cells and screen positive clones with G418 for proliferation. Determine the expression of HBsAg by ELISA and extract the genome DNA of the transgenie cells for amplification by PCR. Results The recombinant plasmid pBIBSb was digested into two fragments at lengths of 700 bp and 3 000 bp respectively, which were consistent with the theoretical value. Eight cell strains growing well were screened by G418, of which five could express HBsAg. A 700 bp fragment was amplified from genome DNA of the transgenic ceils. Conclusion An expression system of HBsAg in ginseng ceils was constructed.

关 键 词：基因转化 乙肝病毒表面抗原 人参细胞 抗生素 中医药疗法 细胞培养 

分 类 号：R512.62[医药卫生—内科学]