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《CHINESE JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY》 2000-02
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The Intracellular Production of Active Horseradish Peroxidase in Methyltrophic Yeast Pichia pastoris

JIANG Tai jiao,JI Xin song,ZHANG Ru an,YUAN Zhong yi (Shanghai Institute of Biochemistry,Chinese Academy of Sciences,Shanghai 200031,China)  
To explore a new approach to express HRP in methyltrophic yeast, Pichia pastoris ,the gene coding for mature HRP isozyme C was subcloned into the intracellular expression vector pPIC3.5K.After transformation of pPIC3.5KHRP into GS115,the positive recombinant P.pastoris strains were screened and obtained by PCR and induced by methanol.Western blot analysis showed that the target protein(about 38 kD)was recognized by polyclonal antibodies against native HRP.This demonstrated that the active horseradish peroxidase was expressed intracellularly in P.pastoris .The intracellular components of positive recombinant P.pastoris strains showed much higher activity of peroxidase than that of recombinant P.pastoris strain only integrated with pPIC3.5K,and the activity of peroxidase was little affected by the addition of heme and CaCl 2 during induction.
【CateGory Index】: Q55
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