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2007•Acta Academiae Medicinae JiangxiRequires access

Recombinant Vector and Screening Multicopy of Human IL-18 in Pichia Pastoris

Song Fang

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Abstract

Objective To study recombinant vector and screen the highest resistance transforming of mature human IL-18(mhIL-18) which can be expressed in high level aod purified effectively in pichia pastoris.Methods The gene of mhIL-18-Intein was amplified by SOEing and asymmetry PCR. And the recombinant expression plasmid pPIC9IL-18-Intein and pPIC9KIL-18-Intein were constructed. The transform containing highest resistance of pichia pastoris were selected with increasing concentration of G418.Results The recombinant vector was identified by transformation,PCR, restriction enzymes digestion analysis and DNA sequencing. The recombinant pichia pastoris was selected by G418.Conclusion The recombinant vector pPIC9KIL-18-Intein is constructed and the highest resistance of pichia pastoris of mhIL-18 is obtained successfully.

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What this paper is about

Objective To study recombinant vector and screen the highest resistance transforming of mature human IL-18(mhIL-18) which can be expressed in high level aod purified effectively in pichia pastoris.Methods The gene of mhIL-18-Intein was amplified by SOEing and asymmetry PCR. And the recombinant expression plasmid pPIC9IL-18-Intein and pPIC9KIL-18-Intein were constructed. The transform containing highest resistance of pichia pastoris were selected with increasing concentration of G418.Results The recombinant vector was identified by transformation,PCR, restriction enzymes digestion analysis and DNA sequencing. The recombinant pichia pastoris was selected by G418.Conclusion The recombinant vector pPIC9KIL-18-Intein is constructed and the highest resistance of pichia pastoris of mhIL-18 is obtained successfully.

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Available abstract

Objective To study recombinant vector and screen the highest resistance transforming of mature human IL-18(mhIL-18) which can be expressed in high level aod purified effectively in pichia pastoris.Methods The gene of mhIL-18-Intein was amplified by SOEing and asymmetry PCR. And the recombinant expression plasmid pPIC9IL-18-Intein and pPIC9KIL-18-Intein were constructed. The transform containing highest resistance of pichia pastoris were selected with increasing concentration of G418.Results The recombinant vector was identified by transformation,PCR, restriction enzymes digestion analysis and DNA sequencing. The recombinant pichia pastoris was selected by G418.Conclusion The recombinant vector pPIC9KIL-18-Intein is constructed and the highest resistance of pichia pastoris of mhIL-18 is obtained successfully.

Key concepts: Pichia pastoris, Intein, Recombinant DNA, Pichia, Transformation (genetics), Biology, Expression vector, Plasmid

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