Construction of human PPAR?_1 LBD fusion protein expression vector and optimization of inducing conditions / 中国药理学通报

ABSTRACT

Aim To obtain high pure hPPAR?1LBD fusion protein.Methods A cDNA encoding ligand binding domain(LBD)of PPAR?1 was amplified by RT-PCR from human fatty tissue and the product was inserted into the downstream of the malE gene in the vector pMAL-p2X,which encoded maltose-binding protein(MBP).The recombinant plasmid containing MBP-PPAR?1 gene was transformed into E.coli.TB1 and the expression conditions of the recombinant strain were optimized.Results The DNA strap of MW(909 bp) was presented after re-combinant plasmid was digested by Hind Ⅲ and BamH Ⅰ.The high efficient expression of MBP-PPAR?1 fusion protein in TB1 cells was observed with 38.54% product of the total cytoplasm proteins when 0.4 mmol?L-1 IPTG and 6 h incubation were taken at 30℃.Conclusion The recombinant vector was successfully constructed.It could high efficiently express hPPAR?1LBD fusion protein in TB1 cells and obtain the hPPAR?1LBD fusion protein with high bioactivity.