Expression and characterization of the hepatitis E virus ORF3 protein in the methylotrophic yeast,Pichia pastoris |
| |
| Institution: | 1. Laboratory of Veterinary Pathology, Universidade Federal de Mato Grosso (UFMT), Av. Fernando Corrêa da Costa, 2367, Boa Esperança, CEP 78069-900, Cuiabá, Mato Grosso, Brazil;2. Veterinary Virology Laboratory, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves, 9090, Agronomia, CEP 91540-000, Porto Alegre, Rio Grande do Sul, Brazil;3. Laboratory of Veterinary Microbiology, Universidade Federal de Mato Grosso (UFMT), Av. Fernando Corrêa da Costa, 2367, Boa Esperança, CEP 78069-900, Cuiabá, Mato Grosso, Brazil;4. Laboratory of Molecular Biology, Universidade Federal de Mato Grosso (UFMT), Av. Fernando Corrêa da Costa, 2367, Boa Esperança, CEP 78069-900, Cuiabá, Mato Grosso, Brazil;1. Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China;2. Scientific Observing and Experimental Station of Veterinary Pharmacology and Veterinary Biotechnology, Ministry of Agriculture, Yangling, Shaanxi, China;3. Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Henan Agriculture University, Zhengzhou, Henan 450002, China;4. Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool L3 5RF, UK;1. Federal Institute for Risk Assessment, Berlin, Germany;2. Friedrich-Loeffler-Institut, Institute for Novel and Emerging Infectious Diseases, Greifswald-Insel Riems, Germany;3. University of Bern, Institute of Ecology and Evolution, Bern, Switzerland;4. Swiss Institute of Bioinformatics, Genopode, Lausanne, Switzerland;5. Clinic for Poultry and Fish Medicine, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine (Vetmeduni Vienna), Vienna, Austria |
| |
| Abstract: | We have used the methylotrophic yeast, Pichia pastoris, to express the open reading frame 3 (ORF3) of the hepatitis E virus (HEV). The ORF3 gene codes for a 123-amino-acid protein that contains highly immunodominant epitopes and is a potentially useful diagnostic and immunoprophylactic antigen. The expressed protein showed positive on immunoblots probed against antibodies raised in rabbit and infected human patient sera. In order to optimize the ORF3 protein expression, we have examined the regulated expression of this protein and characterized it. Unlike its expression in E. coli, the ORF3 protein was present in both the soluble and insoluble fractions of the cell lysate. The expressed protein is not glycosylated and does not undergo any major processing in the host strain. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
