重庆医学2017，Vol.46Issue(17)：2386-2388,3.DOI:10.3969/j.issn.1671-8348.2017.17.026
重组CHO细胞中不同启动子对含MAR表达载体转基因表达的影响
Influence of different promoters on expression of transgene containing MAR expression vector in recombinant CHO cells
摘要
Abstract
Objective To investigate the effect of different promoters on the expression level of transgene containing MAR expression vector in recombinant CHO cells.Methods The CMV promoter and 3-globin MAR were amplified by PCR,then CMV promoter was replaced the SV40 promoter in pCAT1 for constructing the expression vector droved by CMV promoter.The control vectors of pCAT1 and pCAT2 without containing MAR were simultaneously transfected into the CHO cells.Then the stably transfected cell line was screened by G418.The CAT gene expression level was analyzed by ELISA.Results The expression level of CAT enzyme in the cells transfected with MAR-containing vectors was increased compared with the cells transfected by pCATG and pCAT3 vectors without containing MAR,which were increased by 1.75 and 1.25 times respectively(P＜0.05);but CAT enzyme expression level in the pCAT1 transfected cells droved by SV40 promotor with the MAR-containing expression vectors was 1.4 times higher than that in the pCAT2 vector droved by the CMV promoter(P＜0.05).Conclusion MAR can enhance the transgene expression level in stably recombinant CHO cells,and the promoting efficiency of SV40 promoter and MAR combination is superior to that of CMV promoter and MAR combination.关键词
MAR/启动子/转基因表达/CHO细胞Key words
matrix attachment region/promoter/transgene expression/CHO cell分类
生物科学引用本文复制引用
李琴,赵春澎,王小引,孙秋丽,王天云..重组CHO细胞中不同启动子对含MAR表达载体转基因表达的影响[J].重庆医学,2017,46(17):2386-2388,3.基金项目
国家自然科学基金资助项目(31371332). （31371332）
