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2007•Journal of Fujian Normal UniversityRequires access

Expression of Recombinant Human Bone Morphogenetic Protein 4 in Pichia pastoris

Yanding Zhang

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Abstract

Have established techniques to express recombinant human bone morphogenetic proteins 4(rhBMP4) in the methylotrophic yeast Pichia pastoris.The mature domain of hBmp4 was subcloned into the expression vector pPIC9K from recombinant vector pBluescript II KS(+)-hBmp4,and the resulting construct was transformed into Pichia pastoris GS115 for expression by electroporation.It was found with quantity dot blot that rhBMP-4 was expressed with a yield of about 20.13 μg/mL in the supernatant.The expressed rhBMP4 was identified to be a 26 ku monomer protein by SDS-PAGE and Western-blot.

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What this paper is about

Have established techniques to express recombinant human bone morphogenetic proteins 4(rhBMP4) in the methylotrophic yeast Pichia pastoris.The mature domain of hBmp4 was subcloned into the expression vector pPIC9K from recombinant vector pBluescript II KS(+)-hBmp4,and the resulting construct was transformed into Pichia pastoris GS115 for expression by electroporation.It was found with quantity dot blot that rhBMP-4 was expressed with a yield of about 20.13 μg/mL in the supernatant.The expressed rhBMP4 was identified to be a 26 ku monomer protein by SDS-PAGE and Western-blot.

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Available abstract

Have established techniques to express recombinant human bone morphogenetic proteins 4(rhBMP4) in the methylotrophic yeast Pichia pastoris.The mature domain of hBmp4 was subcloned into the expression vector pPIC9K from recombinant vector pBluescript II KS(+)-hBmp4,and the resulting construct was transformed into Pichia pastoris GS115 for expression by electroporation.It was found with quantity dot blot that rhBMP-4 was expressed with a yield of about 20.13 μg/mL in the supernatant.The expressed rhBMP4 was identified to be a 26 ku monomer protein by SDS-PAGE and Western-blot.

Key concepts: Pichia pastoris, Recombinant DNA, Electroporation, Pichia, Molecular biology, Western blot, Expression vector, Yeast

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