Cloning and expression of hFOXP3-Δ E251 in E. coli.

Objective:To clone and express a deletion mutanted human FOXP3 fusion protein,which contains a protein transduction domain from HIV-TAT (PTD-hFOXP3-ΔE251) and lay the groundwork for future researching on the behavior of human FOXP3 in vitro even in vivo.Methods:hFOXP3 cDNA was amplified from total RNA of human peripheral blood mononuclear cells (PBMCs)by reverse transcription-polymerase chain reaction (RT-PCR).Then the expression plasmid named pET28a-PTDhFOXP3-ΔE251 was constructed by genetically cloning the hFOXP3-ΔE251 fragment into the vector pET28a-PTD,which was under control of the T7 promotor.In the end it was expressed in E.coli Rosetta (DE3) induced by isopropyl-β-D-thiogalactopyranoside (IPTG).Results:The result of SDS-PAGE indicated that the fusion protein was highly expressed in E.coli Rosetta (DE3),after induced by 0.5mM IPTG for 8h and the expression product was performed as inclusion body.Conclusion:The expression of PTD-hFOXP3-ΔE251 protein was well done and it should be a potential useful tool for studying the biological functions of hFOXP3.