Rapid expression and purification of the hepatitis delta virus antigen using the methylotropic yeast Pichia pastorisReportar como inadecuado




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BMC Research Notes

, 10:340

First Online: 27 July 2017Received: 01 September 2016Accepted: 26 July 2017

Abstract

ObjectivePatients with dual hepatitis B HBV and hepatitis D HDV virus infection are at an increased risk of progression to liver cirrhosis and hepatocellular carcinoma than patients with a single viral infection. Treatment of viral hepatitis due to dual HBV-HDV infection represents a challenge. Currently there is no vaccine against HDV. Recombinant production of HDV antigen HDAg is the first step towards a potential vaccine candidate and the development of assays for HDV detection.

ResultsThis study demonstrates the expression of one HDAg isoform, S-HDAg, in Pichia pastoris. A recombinant vector carrying a tagged gene encoding S-HDAg under the control of the methanol-inducible promoter AOX1 was designed and integrated into P. pastoris X33. The protein, which was purified using a Ni affinity column and eluted at 100–150 mM imidazole, has potential as a recombinant antigen for further study.

KeywordsHepatitis delta virus HDAg Pichia pastoris Protein expression AbbreviationsBMMYbuffered methanol-complex medium

BMGYbuffered glycerol-complex medium

E. coliEscherichia coli

HDAghepatitis delta antigen

LBLuria Broth

MCSmultiple cloning sites

YPDyeast extract peptone dextrose

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Autor: Stephanie P. Cartwright - Roslyn M. Bill - Bui Tien Sy - Hieu Tran-Van - Hung Minh Nguyen

Fuente: https://link.springer.com/







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