Expression of the Mature Protein Gene of Chicken γ-Interferon and the Detection for the Antiviral Activity of the Purified Expression Product

CAI Mei-hong; CAO Rui-bing; ZHOU Bin; TAN Guo-lei; YANG Song; CHEN Pu-yan

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February 2004

CAI Mei-hong, CAO Rui-bing, ZHOU Bin, TAN Guo-lei, YANG Song and CHEN Pu-yan. Expression of the Mature Protein Gene of Chicken γ-Interferon and the Detection for the Antiviral Activity of the Purified Expression Product[J]. Virologica Sinica, 2004, 19(1): 32-35.

Citation: CAI Mei-hong, CAO Rui-bing, ZHOU Bin, TAN Guo-lei, YANG Song, CHEN Pu-yan. Expression of the Mature Protein Gene of Chicken γ-Interferon and the Detection for the Antiviral Activity of the Purified Expression Product .VIROLOGICA SINICA, 2004, 19(1) : 32-35.

鸡 γ-干扰素成熟蛋白基因的表达及其产物抗病毒活性测定

蔡梅红 ¹ ,
曹瑞兵 ¹ ,
谈国蕾 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.
沈阳农业大学动物医学院, 辽宁沈阳 110161

摘要

以血淋巴细胞提取的总RNA为模板，通过RT-PCR的方法克隆出鸡γ干扰素成熟蛋白的基因，把它与非融合表达载体pRLC相重组。通过对阳性宿主菌的不同时间的诱导摸索出最佳表达时间。把表达产物进行变性，复性，纯化后加入鸡胚成纤维细胞上，用水疱性口炎病毒进行攻毒,测出鸡重组γ干扰素的活性单位为1.0×106U/mL,获得了满意结果。
- 鸡γ干扰素成熟蛋白基因
- , 非融合性表达
- , 病毒抑制
- , 鸡胚成纤维细胞

Expression of the Mature Protein Gene of Chicken γ-Interferon and the Detection for the Antiviral Activity of the Purified Expression Product

1.
1. Key Laboratory of Animal Diseases Diagnosis & Immunology of China’s Department of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
2.
Department of Veterinary medicine,Shenyang Agricultural University, Shenyang 110161, China

Corresponding author: CHEN Pu-yan,

Abstract

Using the total mRNA in the lymphocyte in chicken blood as the template, the mature protein gene of interferon γ-interferon was cloned and amplified through the reverse transcription polymerase chain reaction. The gene was then ligated to pRLC to establish the gene’s non-fusion expressing vector. The best inducing time for the expression of this recombinant protein was tested through inducing the expression of positive selected plasmid with different time span. This expressed product was then denatured and renatured and was added to the culture medium with CEF, which were attacked by the 100 TCID50 VSV. The result showed that the activity of the recombinant protein was up to 1.0×106 U/mg.
- Chicken γ-interferon mature protein gene
- , Non-fusion expression
- , Inhibition of virus
- , CEF

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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 the Mature Protein Gene of Chicken γ-Interferon and the Detection for the Antiviral Activity of the Purified Expression Product

Corresponding author: CHEN Pu-yan,

1. 1. Key Laboratory of Animal Diseases Diagnosis & Immunology of China’s Department of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
2. Department of Veterinary medicine,Shenyang Agricultural University, Shenyang 110161, China

Abstract: Using the total mRNA in the lymphocyte in chicken blood as the template, the mature protein gene of interferon γ-interferon was cloned and amplified through the reverse transcription polymerase chain reaction. The gene was then ligated to pRLC to establish the gene’s non-fusion expressing vector. The best inducing time for the expression of this recombinant protein was tested through inducing the expression of positive selected plasmid with different time span. This expressed product was then denatured and renatured and was added to the culture medium with CEF, which were attacked by the 100 TCID50 VSV. The result showed that the activity of the recombinant protein was up to 1.0×106 U/mg.