Construction and expression of a pEGFP-C2-HDAC2 recombinant plamid

VernacularTitle:重组组蛋白去乙酰化酶2-pEGFP-C2质粒的构建及表达
Author: Hui ZHANG ; Cheng HUANG ; Erbao BIAN ; Bin ZHAO ; Baoming WU ; Changwei LIU ; Xiaoxia CHEN ; Ju LI
Publication Type:Journal Article
Keywords: HDAC2; COS-7; fusion protein; pEGFP-C2; HepG2; GFP
From: Chinese Pharmacological Bulletin 2014;(6):812-815,816
CountryChina
Language:Chinese
Abstract: Aims HDAC2 gene was cloned into pEGFP-C2 vector to explore the efficiency of the plasmid trans-fection in renal fibroblasts COS-7 cells to identify the expression of both mRNA and protein levels and to ob-serve the distribution of the protein. Methods The HDAC2 cDNA was amlified by PCR and cut with the double enzyme Xho I and BamH I, then inserted into the eukaryotic expression vector pEGFP-C2 with T4 en-zyme. The recombinant vector was verified by PCR, restriction enzymes cut and sequencing identification. Then it was transfected into COS-7 cells and the ex-pression of pEGFP-C2-HDAC2 was monitored by fluo-
rescence microscope and PCR. Results Fragments of HDAC2 could be seen after dealt with double diges-tion, and GFP could also be detected in the transfected COS-7 cells. HDAC2 gene expression could be detec-ted by PCR and Western blot. The fusion expression of pEGFP-C2-HDAC2 could be detected by Western blot. Conclusion Eukaryotic expression vector of HDAC2 has been successfully constructed, the fusion expres-sion of HDAC2 and GFP protein can be detected in COS-7 cells.