| 猪流行性腹泻病毒N基因的克隆及原核表达 |
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| 引用本文： | 杨巍,李广兴,孟凡丹,王鑫,任晓峰.猪流行性腹泻病毒N基因的克隆及原核表达[J].东北农业大学学报,2011,42(6):81-85,145. |
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| 作者姓名： | 杨巍 李广兴 孟凡丹 王鑫 任晓峰 |
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| 作者单位： | 东北农业大学动物医学学院,哈尔滨,150030 |
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| 基金项目： | 黑龙江省教育厅“黑龙江省普通高等学校新世纪优秀人才培养计划”项目(1155--NCET--005) |
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| 摘 要： | 从患猪流行性腹泻的病猪粪便中分离了猪流行性腹泻病毒(PEDV)命名为HLJBY，将病毒在Vero细胞上传代，使之适应生长。将扩增的猪流行性腹泻病毒的N基因克隆到原核表达载体PET30a上,构建了重组表达质粒PET-30a-PEDV-N，并在37℃的条件下诱导表达，通过不同时间取样，经SDS-PAGE分析结果表明，该蛋白...
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| 关 键 词： | PEDV N基因 原核表达 N蛋白 |
Cloning and expression of porcine epidemic diarrhea virus N gene |
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| YANG Wei,LI Guangxing,MENG Fandan,WANG Xin,REN Xiaofeng.Cloning and expression of porcine epidemic diarrhea virus N gene[J].Journal of Northeast Agricultural University,2011,42(6):81-85,145. |
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| Authors: | YANG Wei LI Guangxing MENG Fandan WANG Xin REN Xiaofeng |
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| Institution: | YANG Wei,LI Guangxing,MENG Fandan,WANG Xin,REN Xiaofeng(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | The porcine epidemic diarrhea virus(PEDV) that was isolated from the feces of pigs with diarrhea was named HLJBY and was passaged in Vero cell line.The amplified PEDV N gene was cloned into prokaryotic expression vector PET30a,constructing a recombinant expression vector PET-30a-PEDV-N.High level expression of the N protein was analyzed by SDS-PAGE at different time points at 37 ℃ after IPTG induction.The cultured viruses were purified by ultracentrifugation and injected to a rabbit to generate polyclonal antibody.Western-blotting result showed the polyclonal antibody could specifically bind to PEDV N protein,which indicated that the recombinant fusion protein had excellent immunogenicity. |
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| Keywords: | porcine epidemic diarrhea virus N gene prokaryotic expression N protein |
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