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 pTRE2hy-Egr-1 was identified by sequencing. This recombinant vector was transfected into 293Tet-off cells by means of liposome and its expression was examined by Western blotting under the control of deoxycycline.Results:The responsive plasmid pTRE2hyg-Egr-1 verified by sequencing , was capable of expression in 293Tet-off could be controlled by deoxycycline concentration.Conclusion: The responsive plasmid pTRE2hygEgr-1 of Tet-off expression system was constructed successfully, and it can express under the regulation of deoxycycline in the 293Tet-off cells.
收稿日期: 2014-03-04
基金资助:
国家自然科学基金资助项目（31100964）
作者简介: 彭琬昕（1982—），女，安徽合肥人，讲师，主要从事肿瘤细胞生物学研究。
引用本文:
彭琬昕1，2，孟凡力2，3，金洁1，龚爱华1. 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(02): 118-121.
PENG Wan-xin1,2, MENG Fan-li2,3, JIN Jie1, GONG Ai-hua1. Establishment and detection of a Tet-off stable cell line expressing Egr-1 protein regulated by doxycycline. Journal of Jiangsu University(Medicine Edition), 2014, 24(02): 118-121.
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2014, Vol. 24 
