摘要
目的 利用二氢叶酸还原酶缺陷的中国仓鼠卵巢细胞 (CHO dhfr- )和携带二氢叶酸还原酶基因的质粒载体 ,建立高效表达乙型肝炎病毒表面抗原 (HBsAg)的细胞系 ,以提高HBsAg在哺乳动物细胞中的表达产量。方法 将HBsAg基因插入pCI质粒 ,以脂质体方法转染CHO dhfr- 细胞 ,在氨甲喋呤选择压力下 ,筛选HBsAg表达阳性细胞克隆。结果 所获 2 8株单克隆化细胞系 ,18株表达HBsAg,产量高者为 4株。结论 通过HBsAg的高效表达证明 ,二氢叶酸还原酶缺陷细胞株和相应载体是哺乳类动物细胞表达外源基因的有效体系之一。
Objective To set up an efficient expressing system for recombinant hepatitis B virus surface antigen (HBsAg) in dhfr gene negative CHO cell line. Methods HBsAg gene expressing plasmid pCI dhfr S was constructed by integrating HBsAg gene into plasmid pCI which carries dhfr gene. The HBsAg expressing cell line was set up by transfection of plasmid pCI dhfr S into dhfr gene negative CHO cell line in the way of lipofectin. Results Under the selective pressure of MTX, 18 of 28 clonized cell lines expressed HBsAg, 4 of them reached a high titer of 1:32 and protein content 1-3μg/ml. Conclusion In this study, the high level expression of HBsAg demonstrated that the dhfr negative mammalian cell line when recombined with plasmid harboring the corresponding deleted gene can efficiently express the foreign gene. The further steps toward building optimum conditions of the expressing system and the increase of expressed product are under study.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2001年第2期169-170,共2页
Chinese Journal of Experimental and Clinical Virology
