江苏大学学报（医学版）2014，Vol.24Issue(2)：118-121,4.DOI:10.13312/j.issn.1671-7783.y140046
Tet-off诱导表达Egr-1 HEK293稳定细胞株的建立和鉴定
Establishment and detection of a Tet-off stable cell line expressing Egr-1 protein regulated by doxycycline
摘要
Abstract
Objective:To construct the responsive plasmid of pTRE2hyg-Egr-1 Tet-off gene expression system and examine its expression.Methods:PCR was performed to obtain the cDNA of early growth response-1 (Egr-1) which was inserted into the responsive plasmid PTRE2hyg.DNA sequencing was performed after the recombinant of responsive plasmid pTRE2hyg-Egr-1 was identified by sequencing.This recombinant vector was transfected into 293Tet-offcells by means of liposome and its expression was examined by Western blotting under the control of deoxycycline.Results:The responsive plasmid pTRE2hygEgr-1 verified by sequencing,was capable of expression in 293Tet-offcould be controlled by deoxycycline concentration.Conclusion:The responsive plasmid pTRE2hyg-Egr-1 of Tet-off expression system was constructed successfully,and it can express under the regulation of deoxycycline in the 293Tet-offcells.关键词
Tet-off调控系统/早期生长反应蛋白1/稳定细胞株/多西环素Key words
Tet-off inducible gene expression system/ early growth response-1/ stable cell line/ doxycycline分类
医药卫生引用本文复制引用
彭琬昕,孟凡力,金洁,龚爱华..Tet-off诱导表达Egr-1 HEK293稳定细胞株的建立和鉴定[J].江苏大学学报（医学版）,2014,24(2):118-121,4.基金项目
国家自然科学基金资助项目(31100964) （31100964）
