Expression of Avian Infectious Bronchitis Virus E Protein in E.coli

ZHOU Yu-chang; LIU Sheng-wang*; KONG Xian-gang

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February 2006

ZHOU Yu-chang, LIU Sheng-wang* and KONG Xian-gang. Expression of Avian Infectious Bronchitis Virus E Protein in E.coli[J]. Virologica Sinica, 2006, 21(1): 78-80.

Citation: ZHOU Yu-chang, LIU Sheng-wang*, KONG Xian-gang. Expression of Avian Infectious Bronchitis Virus E Protein in E.coli .VIROLOGICA SINICA, 2006, 21(1) : 78-80.

鸡传染性支气管炎病毒E蛋白基因的表达

1.
中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室，黑龙江哈尔滨 150001

摘要
- 传染性支气管炎病毒
- , 重组E蛋白
- , 兔抗IBV E蛋白多克隆血清

Expression of Avian Infectious Bronchitis Virus E Protein in E.coli

1.
National Key Laboratory of Veterinary Biotechnology, Institute of Harbin Veterinary Medicine，Chinese Academy of Agricultural Science, Harbin 150001,China

Abstract

Reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the Small Envelope (E) gene of avian Infectious bronchitis virus (IBV) LX4 strain and the gene was cloned into the pMD18-T vector. By digestion with restriction enzymes SalⅠand BamHⅠ, the E gene was subcloned into pET-30a vector to construct recombinant plasmid pET-30a-E. The recombinant plasmid was transformed into E. coli BL21(DE3) and induced with IPTG. It was demonstrated by SDS-PAGE that a protein of 14kDa, which was comparable in size to the native E protein, was expressed in E. coli. The 14KDa protein was purified and used to prepare the rabbit antiserum against IBV E protein. The result showed that the antibody could react with purified E protein in Western blotting.
- Infectious bronchitis virus (IBV)
- , Recombinant E Protein
- , Rabbit Antiserum

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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

Expression of Avian Infectious Bronchitis Virus E Protein in E.coli

1. National Key Laboratory of Veterinary Biotechnology, Institute of Harbin Veterinary Medicine，Chinese Academy of Agricultural Science, Harbin 150001,China

Abstract: Reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the Small Envelope (E) gene of avian Infectious bronchitis virus (IBV) LX4 strain and the gene was cloned into the pMD18-T vector. By digestion with restriction enzymes SalⅠand BamHⅠ, the E gene was subcloned into pET-30a vector to construct recombinant plasmid pET-30a-E. The recombinant plasmid was transformed into E. coli BL21(DE3) and induced with IPTG. It was demonstrated by SDS-PAGE that a protein of 14kDa, which was comparable in size to the native E protein, was expressed in E. coli. The 14KDa protein was purified and used to prepare the rabbit antiserum against IBV E protein. The result showed that the antibody could react with purified E protein in Western blotting.