Avian influenza A/H5N1 neuraminidase expressed in yeast with a functional head domain
| Autor: | Anan Jongkaewwattana, Yongyuth Yuthavong, L. Eurwilaichitr, Penchit Chitnumsub, Katewadee Boonyapakron, Asawin Wanitchang, Suganya Yongkiettrakul, Ubolsree Leartsakulpanich |
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| Rok vydání: | 2009 |
| Předmět: |
Genes
Viral Genetic Vectors Molecular Sequence Data Orthomyxoviridae Gene Expression Neuraminidase Avian influenza virus medicine.disease_cause Article Pichia Pichia pastoris law.invention Transformation Genetic law Virology Influenza A virus medicine Amino Acid Sequence Glycoproteins chemistry.chemical_classification Influenza A Virus H5N1 Subtype biology H5N1 Neuraminidase (NA) biology.organism_classification Recombinant Proteins Kinetics Enzyme chemistry Biochemistry biology.protein Recombinant DNA Heterologous expression Glycoprotein |
| Zdroj: | Journal of Virological Methods |
| ISSN: | 0166-0934 |
| DOI: | 10.1016/j.jviromet.2008.10.025 |
| Popis: | The study reports heterologous expression in Pichia pastoris of active neuraminidase derived from avian influenza virus A/Viet Nam/DT-036/2005(H5N1). A gene encoding the neuraminidase N1 head domain (residues 63-449) was fused directly in-frame with the Saccharomyces cerevisiae alpha-factor secretion signal in pPICZ(A vector. Recombinant N1 neuraminidase was expressed in P. pastoris as a 72kDa secreted, soluble protein. Glycopeptidase F treatment generated a 45kDa product, indicating that the secreted recombinant N1 neuraminidase is an N-linked glycoprotein. Kinetic studies and inhibition tests with oseltamivir carboxylate demonstrated that the recombinant N1 neuraminidase has similar K(m) and K(i) values to those of the viral N1 neuraminidase. This yeast-based heterologous expression system provided functionally active recombinant N1 neuraminidase that should be useful in anti-influenza drug screening, and also as a potential protein-based vaccine. |
| Databáze: | OpenAIRE |
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