摘要
目的:进一步考察不同G418维持用量与rhGM-CSF表达水平相关性。方法:第2期历时3个月(第4~6个月),本室构建的CHO-rhGM-CSF稳转克隆,在不同G418维持用量(0、300μg/ml、600μg/ml)下传25代左右,各批次传代上清用依赖GM-CSF生长的TF-1细胞作MTT活性测定。结果:无G418选择维持,CHO稳转细胞株表达和分泌hGM-CSF的水平从第4个月开始下降,再次加G418(600μg/ml)后,hGM-CSF的表达水平迅速回升,2周后至正常。结论:Neo抗性基因作为遗传选择标志进入真核细胞,上述结果对于选择G418用量以维持宿主细胞表达分泌目标蛋白具有一定参考价值。
Objective:To a further observa the correlation between the different G418 selection pressure and the rhGM CSF expression level.Methods:During the second term(from the fourth to the sixth month), the stably transfected CHO rhGM CSF clones,constructed by our research department,were cultured under different selection pressure of G418(0,300μg/ml,600μg/ml) for three monthes.The GM CSF activity of culture supernatant of clones was assayed by MTT method with the hGM CSF dependent cell line of TF1.Results:Without G418 selection pressure,the level of rhGM CSF expression and secretion by the clones began descending from the fourth month,whereas the level could rise and return to normal after two weeks when adding G418(600μg/ml) once more.Conclusion:When neo gene as a selective marker came into eucaryotic cells,the above mentioned results may be have a certain reference value for how to select G418 pressure to maintain the expression and secretion of objective proteins by the host cells.
出处
《中国输血杂志》
CAS
CSCD
北大核心
1998年第4期176-178,共3页
Chinese Journal of Blood Transfusion