Construction of Recombinant Adenovirus Containing Pocine IFN-α genes and Activity of Anti-Swine Foot- and-mouth Disease Virus

DU Yi-Jun; JIANG Ping; LI YU-Feng; WANG Xian-Wei

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August 2006

DU Yi-Jun, JIANG Ping, LI YU-Feng and WANG Xian-Wei. Construction of Recombinant Adenovirus Containing Pocine IFN-α genes and Activity of Anti-Swine Foot- and-mouth Disease Virus[J]. Virologica Sinica, 2006, 21(4): 390-393.

Citation: DU Yi-Jun, JIANG Ping, LI YU-Feng, WANG Xian-Wei. Construction of Recombinant Adenovirus Containing Pocine IFN-α genes and Activity of Anti-Swine Foot- and-mouth Disease Virus .VIROLOGICA SINICA, 2006, 21(4) : 390-393.

猪α干扰素重组腺病毒的构建及其抗口蹄疫病毒活性研究

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

通讯作者： 姜平, ;

摘要

本研究利用RT-PCR方法扩增猪α干扰素成熟蛋白基因后构建了重组腺病毒质粒pAd-poIFN-α，经PacI酶切后转染HEK-293A细胞，3次噬斑纯化后获得了重组腺病毒rAd-poIFN-α。该重组腺病毒于HEK-293A细胞连续传代至20代滴度稳定，为107 TCID50/mL。RT-PCR检测证明目的基因在mRNA水平上可有效表达；在PK-15细胞上可以检测到较强的抗猪口蹄疫病毒活性，从而为研究猪口蹄疫免疫防治新技术奠定了重要基础。
- 猪α干扰素
- , 重组腺病毒
- , 口蹄疫病毒

Construction of Recombinant Adenovirus Containing Pocine IFN-α genes and Activity of Anti-Swine Foot- and-mouth Disease Virus

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

Corresponding author: JIANG Ping,

Abstract

Pocine interferon α mature protein genes were amplified by reverse transcription polymerase chain reaction (RT-PCR), and the recombinant replication-defective human adenovius serotype 5 plasmid pAd-poIFN-α was constructed. When the recombinant plasmid pAd-poIFN-α was linearized with PacI, and then transferred into HEK-293A cells, the virus plaque was isolated and purified in HEK-293A cells by three of plaque purification passages. This recombinant adenovirus rAd-poIFN-α could be stably passaged in HEK-293A cells and generated titres in the order of 107 TCID50/mL. The mRNA of the transgene in this recombinant adenovirus was detected by RT-PCR. As a result, the adenovirus exhibited high anti-swine FMDV activity in PK-15 cells and could be further studied in the control strategy of swine FMDV.
- Pocine interferon α
- , Recombinant adenovirus
- , Foot-and-mouth disease virus (FMDV)

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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 of Recombinant Adenovirus Containing Pocine IFN-α genes and Activity of Anti-Swine Foot- and-mouth Disease Virus

Corresponding author: JIANG Ping,

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

Abstract: Pocine interferon α mature protein genes were amplified by reverse transcription polymerase chain reaction (RT-PCR), and the recombinant replication-defective human adenovius serotype 5 plasmid pAd-poIFN-α was constructed. When the recombinant plasmid pAd-poIFN-α was linearized with PacI, and then transferred into HEK-293A cells, the virus plaque was isolated and purified in HEK-293A cells by three of plaque purification passages. This recombinant adenovirus rAd-poIFN-α could be stably passaged in HEK-293A cells and generated titres in the order of 107 TCID50/mL. The mRNA of the transgene in this recombinant adenovirus was detected by RT-PCR. As a result, the adenovirus exhibited high anti-swine FMDV activity in PK-15 cells and could be further studied in the control strategy of swine FMDV.