Construction and Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene

WANG Hai-zhen; Yang Song; SU Xiao-yun; CAO Rui-bing; CHEN Pu-yan

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April 2004

WANG Hai-zhen, Yang Song, SU Xiao-yun, CAO Rui-bing and CHEN Pu-yan. Construction and Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene[J]. Virologica Sinica, 2004, 19(2): 171-173.

Citation: WANG Hai-zhen, Yang Song, SU Xiao-yun, CAO Rui-bing, CHEN Pu-yan. Construction and Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene .VIROLOGICA SINICA, 2004, 19(2) : 171-173.

含双拷贝CSFV E2基因A和D片段原核表达载体的构建及表达*

王海震 ¹ ,
苏小运 ^1,2 ,
曹瑞兵 ¹ ,
陈溥言 ¹ ,

1.
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
2.
沈阳农业大学畜牧兽医学院，辽宁沈阳 100161

摘要
- 猪瘟病毒石门株
- , E2基因
- , 原核串联表达

Construction and Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene

1.
1. Key Lab of Animal Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
2.
College of Animal Science and Veterinary Medicine of Shenyang Agricultural University, Shenyang 110161,China

Corresponding author: CHEN Pu-yan,

Abstract

Full E2 gene fragment of Classical swine fever virus (CSFV)shimen strain was amplified by RT-PCR method, then a specific fraction -the A, D antigenic domains- , which encodes protein with good antigenicity and suitable for being expressed in E. coli with high level, was amplified respectively by 2 different pair of primers. Then the 2 amplified fragments were tandemly linked and inserted into prokaryotic expressing vector pET-32a to obtain the plasmid pET-2e. The SDS-PAGE assay showed that although the proteins were present in the form of inclusion body, the linked proteins were expressed from plasmid pET-2e as expected, and can be expressed with high level when induced with IPTG.Western-blotting showed good antigenicity of the target protein.
- Classical swine fever virus (CSFV)
- , E2 gene
- , Clone and expression

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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 Expression of the Prokaryotic Expressing Vector Containing Tandemly Linked 2 Copies of the A and D Antigenic Domains of the CSFV E2 Gene

Corresponding author: CHEN Pu-yan,

1. 1. Key Lab of Animal Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
2. College of Animal Science and Veterinary Medicine of Shenyang Agricultural University, Shenyang 110161,China

Abstract: Full E2 gene fragment of Classical swine fever virus (CSFV)shimen strain was amplified by RT-PCR method, then a specific fraction -the A, D antigenic domains- , which encodes protein with good antigenicity and suitable for being expressed in E. coli with high level, was amplified respectively by 2 different pair of primers. Then the 2 amplified fragments were tandemly linked and inserted into prokaryotic expressing vector pET-32a to obtain the plasmid pET-2e. The SDS-PAGE assay showed that although the proteins were present in the form of inclusion body, the linked proteins were expressed from plasmid pET-2e as expected, and can be expressed with high level when induced with IPTG.Western-blotting showed good antigenicity of the target protein.