title: Cloning and expression of envelope protein of Thai genotype I strain KE-093 of Japanese encephalitis virus.
abstract: The purpose of this study was to clone and express envelope (E) gene of Japanese encephalitis virus (JEV) genotype I, Thai strain KE-093. The E gene was amplified by PCR and cloned using the expression vector, pET-15b. Analysis of the insert sequence revealed a point mutation, which was corrected by site directed mutagenesis. The envelope 53 kDa protein expression was generated by in vitro coupled transcription translation system. Heterologous expression in Escherichia coli Rosetta 2 strain, but not in E. coli BL21 (DE3) resulted in 2 immunoreactive bands (13 and 53 kDa) using anti-JEV E protein antibodies, and an additional band (35 kDa) using anti-His antibodies, suggesting that E protein antigenicity is located at the carboxy-terminal region. This is the first report of a successful cloning and heterologous expression of an E gene of JEV genotype I. This should prove useful in the application for diagnostics and vaccine development of JEV genotype I strains.
authors.affiliation: Faculty of Science, Burapha University, Chon Buri, Thailand.
authors.forename: Nisakorn
authors.initials: N
authors.lastname: Witthajitsomboon
authors.forename: Antony
authors.initials: A
authors.lastname: Chen
authors.forename: Sasithorn
authors.initials: S
authors.lastname: Lorroengsil
authors.forename: Matti
authors.initials: M
authors.lastname: Sällberg
authors.forename: Somsak
authors.initials: S
authors.lastname: Pantuwatana
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004664:Encephalitis Virus, Japanese / Q000235:genetics*; D005838:Genotype; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D016415:Sequence Alignment; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins