|
Cloning and Expression of HA2 gene of Avian Influenza A (H5N1) Virus in Pichia pastoris |
|---|---|
| รหัสดีโอไอ | |
| Creator | Prapassorn Channoi, Chaisuree Suphawilai and Ronachai Pratanaphon |
| Title | Cloning and Expression of HA2 gene of Avian Influenza A (H5N1) Virus in Pichia pastoris |
| Publisher | Faculty of Agro-Industry |
| Publication Year | 2561 |
| Journal Title | Food and Applied Bioscience |
| Journal Vol. | 6 |
| Journal No. | 2 |
| Page no. | 106-116 |
| Keyword | Avian influenza, Haemagglutinin, H5N1, HA2, Pichia pastoris |
| URL Website | www.tci-thaijo.org/index.php/fabjournal |
| Website title | www.tci-thaijo.org/index.php/fabjournal/article/view/105681 |
| ISSN | 2286-8615 |
| Abstract | Avian influenza or bird flu is a highly infectious viral disease and is one of the most major concerns to both poultry industry and human health. The haemagglutinin (HA) protein is one of the markers for detection of influenza viral infection. There is no efficient protection and therapy for the disease, hence, rapid and accurate detection of avian influenza virus infection is an important tool to control outbreaks.In this study, the HA2 gene was amplified and cloned into pPICZA expression vector. The recombinant plasmid was verified by PCR, restriction analysis and nucleotide sequencing. After transformed into Pichia pastoris, the integration of HA2 gene into host genome was confirmed. The expression of HA2 was performedandthe protein was internally expressed as tagged fusion protein. The recombinantHA2 protein was extracted and purified from cell lysate using nickel affinity chromatography under native condition.A single band of 35-40 kDa was observed bySDS-PAGE. Western blot analysisrevealed that the protein could be reacted with His DetectorTMand anti myc antibody, indicating that this was protein of interest. The HA2 recombinant protein could react with serum sample from patients recovered from avian influenza infection who had their hemaglutination inhibition (HI)titer over 1:40, while it did not react with the non-exposed individual serum. Therecombinant HA2 expressed by P. pastoriscould be useful as a potential antigen for epidemiologicalstudy of highly pathogenic avian influenza (HPAI) infection. |
