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| Establishment and detection of a Tet-off stable cell line expressing Egr-1 protein regulated by doxycycline |
| PENG Wan-xin1,2, MENG Fan-li2,3, JIN Jie1, GONG Ai-hua1 |
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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.
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Received: 04 March 2014
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2014, Vol. 24 
