title: Antigenic validation of recombinant hemagglutinin-neuraminidase protein of Newcastle disease virus expressed in Saccharomyces cerevisiae.
abstract: The outer membrane glycoprotein, hemagglutinin-neuraminidase (HN) of Newcastle disease virus (NDV) is important for virus infection and subsequent immune response by host, and offers target for development of recombinant antigen-based immunoassays and subunit vaccines. In this study, the expression of HN protein of NDV is attempted in yeast expression system. Yeast offers eukaryotic environment for protein processing and posttranslational modifications like glycosylation, in addition to higher growth rate and easy genetic manipulation. Saccharomyces cerevisiae was found to be better expression system for HN protein than Pichia pastoris as determined by codon usage analysis. The complete coding  sequence of HN gene was amplified with the histidine tag, cloned in pESC-URA under GAL10 promotor and transformed in Saccharomyces cerevisiae. The recombinant HN (rHN) protein was characterized by western blot, showing glycosylation heterogeneity as observed with other eukaryotic expression systems. The recombinant protein was purified by affinity column purification. The protein could be further used as subunit vaccine.
authors.forename: S A
authors.initials: SA
authors.lastname: Khulape
authors.forename: H K
authors.initials: HK
authors.lastname: Maity
authors.forename: D C
authors.initials: DC
authors.lastname: Pathak
authors.forename: C Madhan
authors.initials: CM
authors.lastname: Mohan
authors.forename: S
authors.initials: S
authors.lastname: Dey
mesh_terms: D015707:HN Protein / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D009522:Newcastle disease virus / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics*
chemical_list: D015707:HN Protein; D011994:Recombinant Proteins
keywords: Newcastle disease virus; glycoprotein; codon usage; yeast expression.