Expression, Purification And Function Research Of Human LXRα

Purpose:The biological activity of LXRα binding to Hydroxy cholesterol were studied through expression and purification of the recombinant proteins of LXRα in Pichia pastoris(P. pastoris) expression system.Methods:The target gene of LXRα were amplified by PCR methods with the total RNA of THP-1 as template. The recombinant expression vector pPICZαALXRα was constructed and transformed into P. pastoris X-33 cells.The recombinant proteins was induced by methanol and purified by Ni ion affinity column. The binding activity of the recombinant proteins was identified by ELISA.Results:The target DNA of LXRα inserted into eukaryotic expression vector pPICZαA to yield pPICZαA- LXRα. The recombinant protein was highly expressed in P. pastoris after methanol induction. The recombinant LXRα protein expressed in P.pastoris(P.rLXRα) was purified with high purification by Ni ion affinity chromatography. Furthermore, ELISA analysis showed that P.rLXRα have the binding activity to Hydroxy cholesterol as natural LXRα does.Conclusion : The eukaryotic expression vector pPICZαA-LXRα were successfully constructed. The P.rLXRα with high purification was obtained through secretion expressed in P.pastoris expression system. Our results showed that P.r LXRα have the binding activity to Hydroxy cholesterol as natural LXRα does.The work layed a foundation for further research on the Drug Design.