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《Journal of Hubei University(Natural Science)》 2018-01
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The expression of optimized and synthesized organic phosphorus hydrolase opd p gene in Pichia pastoris

WANG Ying;HUANG Can;SUN Qimeng;MA Lixin;YAN Hong;College of Life Sciences,Hubei University;  
In this study,code-optimized organophosphorus hydrolase gene( opd) and bio-brick method were used to construct opd multi-copy vector to increase the expression in Pichia pastoris. The original opd gene was designed to synthesize the new organophosphatase gene according to Pichia pastoris and six His-Tags were added at the C-terminus. The total length of the synthesized opd p gene was 1 149 bp( base pair). The recombinant plasmids of p HBM905 BDM-opd p, p HBM905 BDM-opd p-2 C, p HBM905 BDM-opd p-3 C,p HBM905 BDM-opd p-4 C were constructed and transformed into Pichia pastoris GS115 competent cells. The results showed that OPD P protein was secreted and expressed under the AOX1( alcohol oxidase gene)promoter control,which was induced by methanol. Western Blot showed that the target protein was expressed in the four transformed strains,and the expression level of 2 copies was improved compared with 1 copy,was increased to 0. 2 U/m L. But the expression level did not increase with the increase of copy number.
【Fund】: 国家自然科学基金(31300074)资助
【CateGory Index】: Q55;Q78
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