High Level Secretionary Expression of the ORF2 Gene of Porcine Circovirus in Yeast

ZHANG Xiao-yong; ZhENG Qi-sheng; WEI Xue-tao; TAN Guo-lei; REN Xue-reng; CAO Rui-bing; CHEN De-sheng; CHEN Pu-yan*

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April 2005

ZHANG Xiao-yong, ZhENG Qi-sheng, WEI Xue-tao, TAN Guo-lei, REN Xue-reng, CAO Rui-bing, CHEN De-sheng and CHEN Pu-yan*. High Level Secretionary Expression of the ORF2 Gene of Porcine Circovirus in Yeast[J]. Virologica Sinica, 2005, 20(2): 125-129.

Citation: ZHANG Xiao-yong, ZhENG Qi-sheng, WEI Xue-tao, TAN Guo-lei, REN Xue-reng, CAO Rui-bing, CHEN De-sheng, CHEN Pu-yan*. High Level Secretionary Expression of the ORF2 Gene of Porcine Circovirus in Yeast .VIROLOGICA SINICA, 2005, 20(2) : 125-129.

猪圆环病毒II型ORF2基因在酵母中的高效分泌表达

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

摘要

为了建立一种简便的检测猪圆环病毒2型(PCV2)的方法,本实验将PCV2 的ORF2 基因片段整合到巴斯德毕赤酵母(Pichia pastoris)菌株X 33染色体上,构建了X 33(pPICZa ORF2)重组工程菌。经甲醇诱导后,成功的表达出ORF2基因片段。经过Bradford 蛋白质总含量测定和凝胶薄层扫描结果表明,表达产物占重组工程菌培养上清总蛋白的58%,表达量可达47mg/ L。间接ELISA结果初步表明重组表达产物具有良好的抗原性,能够有效地区分PCV2型病毒标准阳性与阴性血清。
- 猪圆环病毒
- , 巴斯德毕赤酵母
- , 分泌表达

High Level Secretionary Expression of the ORF2 Gene of Porcine Circovirus in Yeast

Abstract

In order to establish a convenient method to detect Porcine Circovirus(pcv2),the ORF2 gene of pcv2 was intergraded into the genome of methylotropic yeast Pichia pastoris to construct an engineering yeast.The ORF2 fragment was successfully secreted into the medium after induced with methanol.Gel thin layer scanning and Bradford total protein analysis results showed that the amount of the expressed product was about 47mg/ L,Compromising 58% of total culture supernatant protein in X-33/pPICZa-ORF2.Indirect ELISA showed the expressed product can be a good antigen and can differentiate the standard positive and negative serum of pcv2 effctively.
- Porcine circovirus Secreting expression Pichia pastoris

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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

High Level Secretionary Expression of the ORF2 Gene of Porcine Circovirus in Yeast

Abstract: In order to establish a convenient method to detect Porcine Circovirus(pcv2),the ORF2 gene of pcv2 was intergraded into the genome of methylotropic yeast Pichia pastoris to construct an engineering yeast.The ORF2 fragment was successfully secreted into the medium after induced with methanol.Gel thin layer scanning and Bradford total protein analysis results showed that the amount of the expressed product was about 47mg/ L,Compromising 58% of total culture supernatant protein in X-33/pPICZa-ORF2.Indirect ELISA showed the expressed product can be a good antigen and can differentiate the standard positive and negative serum of pcv2 effctively.