[Construction and expression of a fusion protein containing extracellular domain of human Delta-like1 and Fc fragment of human IgG1 in the Pichia pastoris].

AIM: To construct a Pichia pastoris expression vector PIC- hDll1(ext)-Fc, and to express the fusion protein containing the extracellular domain of human Delta-like1 and the Fc fragment of human IgG1 fusion gene in Pichia pastoris. METHODS: The extracellular domain of human Delta-like1 gene was amplified from pEF-BOSneo-hdll1(ext)-Fc by PCR. The expression vector was constructed by DNA recombination. The constructed plasmid was transformed into yeast GS115 by electroporation. The recombinant transformants with a high copy number of the plasmid were selected by using MD plate and G418. The expression of protein was induced by addition of methanol.Then analyzed protein expression by SDS-PAGE and Western blot. RESULTS: The extracellular domain of human Delta-like1 gene was effectively amplified. The DNA sequencing result showed that the constructed plasmid containing hDll(ext)-Fc fusion gene was the same as designed. The fusion protein was successfully expressed in Pichia pastoris. CONCLUSION: The hDll1(ext) gene has been successfully cloned and expressed in the form of Fc fusion protein, which provides a new fusion protein for further experiments.