Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2

Cap protein

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February 2006

Cap protein, Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2[J]. Virologica Sinica, 2006, 21(1): 29-33.

Citation: Cap protein, . Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2 .VIROLOGICA SINICA, 2006, 21(1) : 29-33.

猪圆环病毒Ⅱ型Cap蛋白重组腺病毒的构建与鉴定

1.
南京农业大学农业部动物疫病诊断与免疫重点开放实验室，江苏南京 210095

摘要

猪圆环病毒Ⅱ型（PCV2）是近年来新发现的引起仔猪多系统衰弱综合症的必需病原，含有两个主要的阅读框ORF1和ORF2，分别编码复制相关蛋白（Rep）和核衣壳蛋白（Cap），其中Cap含有病毒主要抗原表位，是研究PCV2基因工程苗的主要候选目的基因。本研究利用PCR技术将Cap蛋白基因克隆入人5型腺病毒穿梭载体（pShuttle-CMV），与腺病毒骨架载体(pAdEasyTM)共转化大肠杆菌BJ5183进行同源重组并转染HEK-293A细胞，经多次亚克隆获得了重组腺病毒rAd-Cap，测定其TCID50为1013.7/mL。用RT-PCR、间接ELISA、Western blot和IPMA等方法证明了Cap蛋白在腺病毒中获得表达。该研究为发展PCV2的重组腺病毒基因工程疫苗奠定了基础。
- Ⅱ型猪圆环病毒
- , Cap蛋白
- , 重组
- , 腺病毒
- , 构建

Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2

Cap protein ,
,

1.
Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095,China

Abstract

Porcine circovirus 2 (PCV2) has been implicated as the etiological agent of a new disease, named postweaning multisystemic wasting syndrome (PMWS). PCV2 contains two major open reading frames (ORFs): ORF1 and ORF2, encoding replication-associated (Rep) protein and capsid (Cap) protein, respectively. The cap protein is a candidate 2 for recombinant PCV vaccine, because it possesses several potential antigen epitopes. In the study, the cap gene was amplified by PCR, and cloned into the transfer vector pShuttle-CMV subsequentially. The recombinant plasmid and bone vector pAdEasy-1 were cotransformed into BJ5183 to generate the recombinant plasmid rAd-ORF2 pAd-Cap. The recombinant adenovirus (rAd-Cap) was generated by transfection and plaque purification in in 293 cells. The transcription and expression of Cap protein in rAd-Cap infected 293 cells were confirmed by RT-PCR、indirect-ELISA, Western blot and IPMA. The study laid the foundation for development of the recombinant vaccine against PCV2.
- PCV2
- , Cap protein
- , Recombinant
- , Adenovirus
- , Construction

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction and Identification of Recombinant Adenoviruses Containingthe Cap Protein of Porcine Circovirus 2

Cap protein ,
,

1. Key Laboratory of Animal Diseases Diagnostic and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095,China

Abstract: Porcine circovirus 2 (PCV2) has been implicated as the etiological agent of a new disease, named postweaning multisystemic wasting syndrome (PMWS). PCV2 contains two major open reading frames (ORFs): ORF1 and ORF2, encoding replication-associated (Rep) protein and capsid (Cap) protein, respectively. The cap protein is a candidate 2 for recombinant PCV vaccine, because it possesses several potential antigen epitopes. In the study, the cap gene was amplified by PCR, and cloned into the transfer vector pShuttle-CMV subsequentially. The recombinant plasmid and bone vector pAdEasy-1 were cotransformed into BJ5183 to generate the recombinant plasmid rAd-ORF2 pAd-Cap. The recombinant adenovirus (rAd-Cap) was generated by transfection and plaque purification in in 293 cells. The transcription and expression of Cap protein in rAd-Cap infected 293 cells were confirmed by RT-PCR、indirect-ELISA, Western blot and IPMA. The study laid the foundation for development of the recombinant vaccine against PCV2.