| 摘要: |
| [目的]构建表达PRRSV GP5和M蛋白、且二者通过口蹄疫病毒(FMDV)2A序列连接的重组腺病毒,以期实现2个蛋白的同时表达,发挥GP5蛋白的病毒中和优势和M蛋白的细胞免疫优势作用.[方法]利用RT-PCR、间接免疫荧光和Western blot等方法,对获得的重组腺病毒(rAd-GP5-2A-M)进行检测.[结果]检测结果均证明,GP5-2A-M蛋白不仅获得了正确表达,而且能自我裂解为GP5和M蛋白.[结论]在GP5和M蛋白之间插入具有自我裂解功能的FMDV 2A策略是切实可行的,并为PRRSV多个蛋白的共表达奠定了基础,也为其他病毒基因工程疫苗的研制提供了全新思路. |
| 关键词: 猪繁殖与呼吸综合征病毒 口蹄疫病毒 2A蛋白酶 |
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| 基金项目:浙江省自然科学基金项目（Y305031;Y307124）；浙江省科技厅重点项目（2006C22047）；浙江省重大科技专项（2007C12G4010009）；嘉兴科技局项目（2008AY1001） |
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| Construction and identification of recombinant adenovirus containing FMDV 2A sequence,expressing GP5 and M protein of porcine reproduction and respiratory syndrome virus |
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| Abstract: |
| 【Objective】 Based on the self-cleaving ability of foot-and-mouth disease virus (FMDV) 2A proteinase,we ligated GP5 and M coding region of porcine reproduction and respiratory syndrome virus (PRRSV) with the short peptide,and put them in the expression box of adenovirus vector.Through this new strategy,GP5 and M not only can be expressed at the same time,but also can be cleaved into two independent proteins,which will be helpful to play the neutralizing action of GP5 protein and the cell mediated immunity of M protein.【Method】 The recombinant adenovirus(rAd-GP5-2A-M) was identified by RT-PCR,IFA and Western blot methods etc.【Result】 This kind of recombinant adenovirus （rAd-GP5-2A-M ）was constructed successfully.【Conclusion】 It showed that it is possible to develop one multi-gene engineering vaccine against PRRS utilizing FMDV 2A proteinase,which also can provide a novel strategy for developing other viral disease vaccine. |
| Key words: porcine reproduction and respiratory syndrome virus foot-and-mouth disease virus 2A proteinase |
