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The high expression of HBsAg in Pichia pastoris

HE Xiao-wei

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Abstract

Objective: To secrete the Pichia pastoris strain which has the highest expression of HBsAg. Methods: The HBV S gene was amplified by PCR and cloned into the expression vector pPICZA. Then the reconstructed plasmid pPICZA-S and pPICZA-SH were transformed into the following Pichia pastoris strains: GS115 X-33、KM71H and SMD1168 through LiCl chemical method. Results: The results indicated that the highest expressed Pichia pastoris strain is GS 115 with expression level of 15.3 mg/L and the best harvest time point is 72 hours after induced, because adding the 0.1% casamino acid into inductive medium can restrict the hydrolysis of desired proteins and enhance their expression. Conclusion: HbsAg analyzed by SDS-PAGE and Western-blot was expressed after induced with methanol.

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What this paper is about

Objective: To secrete the Pichia pastoris strain which has the highest expression of HBsAg. Methods: The HBV S gene was amplified by PCR and cloned into the expression vector pPICZA. Then the reconstructed plasmid pPICZA-S and pPICZA-SH were transformed into the following Pichia pastoris strains: GS115 X-33、KM71H and SMD1168 through LiCl chemical method. Results: The results indicated that the highest expressed Pichia pastoris strain is GS 115 with expression level of 15.3 mg/L and the best harvest time point is 72 hours after induced, because adding the 0.1% casamino acid into inductive medium can restrict the hydrolysis of desired proteins and enhance their expression. Conclusion: HbsAg analyzed by SDS-PAGE and Western-blot was expressed after induced with methanol.

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Available abstract

Objective: To secrete the Pichia pastoris strain which has the highest expression of HBsAg. Methods: The HBV S gene was amplified by PCR and cloned into the expression vector pPICZA. Then the reconstructed plasmid pPICZA-S and pPICZA-SH were transformed into the following Pichia pastoris strains: GS115 X-33、KM71H and SMD1168 through LiCl chemical method. Results: The results indicated that the highest expressed Pichia pastoris strain is GS 115 with expression level of 15.3 mg/L and the best harvest time point is 72 hours after induced, because adding the 0.1% casamino acid into inductive medium can restrict the hydrolysis of desired proteins and enhance their expression. Conclusion: HbsAg analyzed by SDS-PAGE and Western-blot was expressed after induced with methanol.

Key concepts: Pichia pastoris, HBsAg, Pichia, Recombinant DNA, Expression vector, Western blot, Molecular biology, Plasmid

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