| Asia1型FMDV非结构蛋白3A基因的克隆、表达与抗原性鉴定 |
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| 引用本文： | 宋军,高明春,宋鸽,刘丹丹,王君伟.Asia1型FMDV非结构蛋白3A基因的克隆、表达与抗原性鉴定[J].黑龙江畜牧兽医,2012(5):19-21,172. |
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| 作者姓名： | 宋军 高明春 宋鸽 刘丹丹 王君伟 |
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| 作者单位： | 东北农业大学动物医学学院 |
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| 基金项目： | 现代农业产业技术体系建设专项资金项目(CARS-37);黑龙江省科技计划项目(GA06B202) |
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| 摘 要： | 为了研究Asia1型口蹄疫病毒(FMDV)非结构蛋白3A的抗原性,试验对Asia1型口蹄疫病毒非结构蛋白3A基因进行扩增、亚克隆及测序,将3A克隆至表达载体pET-32a(+)中,选取阳性克隆转化Rosetta(DE3)pLysS大肠杆菌感受态细胞,用IPTG诱导表达和纯化3A蛋白,并进行SDS-PAGE鉴定与Western-blot分析。结果表明:在大肠杆菌中成功地表达了3A蛋白,表达的目的蛋白能与Asia1型FMDV阳性血清发生特异性反应。说明非结构蛋白3A具有较好的抗原活性。
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| 关 键 词： | 口蹄疫病毒(FMDV) 非结构蛋白 抗原性鉴定 |
Cloning,prokaryotic expression and antigenicity identification of nonstructural protein 3A of Foot-and-mouth disease virus serotype Asia1 |
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| SONG Jun,GAO Ming-chun,SONG Ge,LIU Dan-dan,WANG Jun-wei.Cloning,prokaryotic expression and antigenicity identification of nonstructural protein 3A of Foot-and-mouth disease virus serotype Asia1[J].Heilongjiang Animal Science And veterinary Medicine,2012(5):19-21,172. |
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| Authors: | SONG Jun GAO Ming-chun SONG Ge LIU Dan-dan WANG Jun-wei |
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| Institution: | (College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | To study the prokaryotic expression and antigenicity identification of nonstructural protein 3A gene from Foot-and-mouth disease virus(FMDV) serotype Asia1,the 3A gene fragments were amplified by PCR and subcloned from the Foot-and-mouth disease virus serotype Asia1.Their expressing plasmids were constructed by inserting the 3A gene fragments into pET-32a vector and then transferred into E.coli Rosetta(DE3)pLysS.It was induced and expressed by IPTG and then made for SDS-PAGE identification and Western-blot detecting analysis.The results showed that the 3A protein was successfully expressed in E.coli and the expressed 3A protein could be specifically reacted with the positive serum of FMDV serotype Asia1.This study may provide the basis for establishing the identification and diagnosis method,which can distinguish the infected animals from the vaccinated animals with the recombinant FMDV nonstructural protein as the antigen. |
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| Keywords: | Foot-and-mouth disease virus(FMDV) non-structural protein antigenicity analysis |
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