頁籤選單縮合
題名 | Expression, Purification, and Characterization of the Infectious Bursal Disease Virus-like Particles Produced by Insect Cells= |
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作者 | 何靜宜; 賴素媛; 李龍湖; 王敏盈; Ho, Jin-yi; Lai, Su-yuan; Lee, Long-huw; Wang, Min-ying; |
期刊 | Journal of the Chinese Chemical Society |
出版日期 | 19991000 |
卷期 | 46:5 1999.10[民88.10] |
頁次 | 頁743-750 |
分類號 | 414.84 |
語文 | eng |
關鍵詞 | Infectious bursal disease virus; Virus-like particles; Baculovirus expression system; |
英文摘要 | The cDNA of the large genomic segment A (ORF A1) of infectious bursal disease virus (IBDV) strain P3009 was synthesized and cloned into the transfer vector pBlueBac4. Following cotransfection and plaque purification, a plaque-pure recombinant baculovirus, vIBDVJ6, was selected to express IBDV capsid proteins. When Sf9 cells were infected with vIBDVJ6, IBDV precursor protein (polypeptide of VP2-VP4-VP3) was expressed and proteolytically processed to give VP2, VP3, and VP4 proteins by a viral protease. These resulting recombinant proteins can self-assemble to form virus-like particles (VLPs). The shape and size of the negatively stained purified IBDV-like particles were demonstrated to be similar to those of the complete P3009 IBDV particles under direct electron microscopic observation. Furthermore, the purified particles were strongly recognized by an anti-VP2 monoclonal antibody as confirmed by an immuno-gold labeling experiment. IBDV-like particles will offer a unique opportunity to create completely non-infectious viral vaccines for use in the vaccination of chickens. The immunologic response in chickens stimulated by IBDV-like particles is currently being evaluated. |
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