2005•Biotechnology(Faisalabad)Requires access
Cloning and Identification of Horseradish Peroxidase Isozyme C Gene in Pichia pastoris
Jinxiang Han
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Abstract
Objective:To clone the Horseradish peroxidase isozyme C(HRPC2) of horseradishfor the expression of the gene.Methods:The HRPC2 gene was amplified with PCR from the total DNA of Horseradish.After introns of HRPC2 were cutted,it was linked into pMD-18Tvector.When it was confirmed by sequencing,the target gene was cut down by restriction endonuclease.Then it was inserted into expression vector pPIC9K of Pichia pastoris ,and the recombinant plasmid pC2EX9K was obtained.The HRPC2 gene was cloned in Pichia pastoris and was identified.Results:The recombinant plasmid pC2EX9K was transformed into Pichia pastoris .Then,the cloned target gene was confirmed by PCR analysis.Conclusion:The HRPC2 gene was cloned successfully using pPIC9K as the vector and using Pichia pastoris as host bateria.