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Comparison of two eukaryon expression systems
Author(s): 
Pages: 46-49
Year: Issue:  1
Journal: CHINESE JOURNAL OF IMMUNOLOGY

Keyword:  真核表达转基因细胞表达效率;
Abstract: 目的:比较目的基因在两种不同表达体系构建的转基因细胞上的表达,选择更好的构建体系.方法:脂质体法将重组真核细胞表达载体pcDNA3/PD-L1导入L929细胞,G418(600 μg/ml)筛选抗性细胞株,同时将重组逆转录病毒载体pGEZ-Term/PD-L1与辅助载体共转染293T细胞,包装病毒颗粒,其上清感染L929细胞后,经Zeocin(500 μg/ml) 筛选抗性细胞.流式细胞仪(FCM)检测目的蛋白在两种细胞膜上的表达.结果:两种体系对目的基因的瞬时表达无明显差别;用pcDNA3构建的转基因细胞膜上目的分子表达稳定性差,表达率低,而用pGEZ-Term构建的则目的蛋白稳定高表达,表达率近100%.结论:两种构建体系均能瞬时表达目的分子,pGEZ-Term表达体系更适于构建长期稳定表达目的基因的转基因细胞.
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