| 猪干扰素α1与胸腺肽α1融合蛋白原核表达系统的建立 |
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| 引用本文： | 张鹏,李明谦,郝林琳,马强,张英,刘松财.猪干扰素α1与胸腺肽α1融合蛋白原核表达系统的建立[J].吉林农业大学学报,2012(4):449-453. |
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| 作者姓名： | 张鹏 李明谦 郝林琳 马强 张英 刘松财 |
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| 作者单位： | 吉林大学畜牧兽医学院 |
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| 基金项目： | 吉林省科技支撑计划重点项目(3D510S086604) |
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| 摘 要： | 依据GenBank已发表的猪干扰素α1(poIFN-α1)成熟肽基因序列,胸腺肽α1(THY-α1)蛋白序列及Escherichia coliB密码子使用频率表,优化并人工合成poIFN-α1和THY-α1基因,利用SOE-PCR法在poIFN-α1和THY-α1之间设计1个13肽Linker。将融合基因克隆至pRSFDue-t 1表达载体,转化BL21(DE3)感受态,37℃培养至菌液OD600为0.6~0.8,IPTG(0.5 mmol/L)诱导,20℃培养12h,SDS-PAGE和Western blot鉴定。结果表明:目的蛋白为菌体内可溶性表达,经强阴离子交换层析和疏水层析纯化获得高纯度poIFNα1-THYα1融合蛋白。
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| 关 键 词： | 猪 干扰素α1 胸腺肽α1 融合蛋白 |
Establishment of Prokaryotic Expression System of Porcine IFNα1-THYα1 Fusion Protein |
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| ZHANG Peng,LI Ming-qian,HAO Lin-lin,MA Qiang,ZHANG Ying,LIU Song-cai.Establishment of Prokaryotic Expression System of Porcine IFNα1-THYα1 Fusion Protein[J].Journal of Jilin Agricultural University,2012(4):449-453. |
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| Authors: | ZHANG Peng LI Ming-qian HAO Lin-lin MA Qiang ZHANG Ying LIU Song-cai |
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| Institution: | College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China |
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| Abstract: | The combined application of interferon and thymosin is significant in antiviral effect.According to the genic sequences of porcine interferon-α1 in GenBank,the protein sequences of thymosin-α1 and the codon bias in Escherichia coli B,the poIFN-α1 and THY-α1 genes were optimized and synthesized.A linker containing 13 amino acids was designed between poIFN-α1 and THY-α1 by SOE-PCR.The poIFNα1-THYα1 fusion gene was cloned into pRSFDuet-1 expression plasmid and transformed into BL21(DE3),and then cultured at 37 ℃ and induced with IPTG(0.5 mmol/L) for 12 hours while its OD600 got up to 0.6—0.8,SDS-PAGE and Western blot results indicated the soluble poIFNα1-THYα1 fusion protein was successfully expressed.The high-purity poIFNα1-THYα1 fusion protein was obtained by ion exchange chromatography and hydrophobic chromatography,which made a important sense of prevention and treatment of porcine viral diseases. |
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| Keywords: | porcine IFN-α1 THY-α1 fusion protein |
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