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Expression of Human IL-12 P40 Subunit in Pichia pastoris

Xinhua Chen, Lin Yang, Chen Quhou, Xunzhang Wang, Qingxin Long

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Abstract

A 940 bp coding fragment of human IL-12 p 40 gene was amplified via PCR technique and then cloned into Pichia pastoris expression vector pPICZαC. After transforming the linearized recombinant expression vector into the yeast strain X\|33,we obtained the engineered yeast strain Pichia pastoris X\|33/ p 40 The p 40 gene was secretly expressed after induction with 0 5% methanol Both the result of SDS\|PAGE and Western\|blotting proved a 44 000 protein band in the supernatant of medium. Gel scanning revealed the expression product accounted for 47% of the total supernatant protein.

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What this paper is about

A 940 bp coding fragment of human IL-12 p 40 gene was amplified via PCR technique and then cloned into Pichia pastoris expression vector pPICZαC. After transforming the linearized recombinant expression vector into the yeast strain X\|33,we obtained the engineered yeast strain Pichia pastoris X\|33/ p 40 The p 40 gene was secretly expressed after induction with 0 5% methanol Both the result of SDS\|PAGE and Western\|blotting proved a 44 000 protein band in the supernatant of medium. Gel scanning revealed the expression product accounted for 47% of the total supernatant protein.

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Available abstract

A 940 bp coding fragment of human IL-12 p 40 gene was amplified via PCR technique and then cloned into Pichia pastoris expression vector pPICZαC. After transforming the linearized recombinant expression vector into the yeast strain X\|33,we obtained the engineered yeast strain Pichia pastoris X\|33/ p 40 The p 40 gene was secretly expressed after induction with 0 5% methanol Both the result of SDS\|PAGE and Western\|blotting proved a 44 000 protein band in the supernatant of medium. Gel scanning revealed the expression product accounted for 47% of the total supernatant protein.

Key concepts: Pichia pastoris, Recombinant DNA, Yeast, Molecular biology, Pichia, Expression vector, Protein subunit, Gene

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