| Asia1型口蹄疫病毒P1基因的原核表达及其抗血清的制备 |
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| 引用本文： | 武刚,王洪梅,刘晓,王立群,于力,仲跻峰,何洪彬.Asia1型口蹄疫病毒P1基因的原核表达及其抗血清的制备[J].农业科学与技术,2010(9):112-114,143. |
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| 作者姓名： | 武刚 王洪梅 刘晓 王立群 于力 仲跻峰 何洪彬 |
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| 作者单位： | [1]山东省农业科学院奶牛研究中心,山东济南250100 [2]东北农业大学生命科学院,黑龙江哈尔滨150030 [3]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150000 |
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| 基金项目： | 国家转基因重大专项（2009ZX08007-006B）;国家自然科学基金（31072160）;山东省科技攻关课题（2009GG20002032）;山东省自然基金（Y2008D20）;兽医生物技术国家重点实验室开放课题基金（SKLVBF200806）. |
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| 摘 要： | 目的]研究Asial型口蹄疫病毒P1基因原核表达及其抗血清的制备。方法]利用基因克隆技术获得Asia1型口蹄疫病毒（FMDV）的P1基因,然后将P1基因重组到pET-32a（＋）质粒中;转化大肠杆菌BL21感受态细胞,经IPTG诱导及蛋白纯化后,进行SDS-PAGE;将重组菌BL21培养物用超声波裂解,对该融合蛋白进行分离纯化后免疫新西兰兔,制备P1蛋白抗血清。结果]获得了重组性阳性克隆;SDS-PAGE结果表明在105kD处出现了目的条带;Western-blot分析表明,抗血清可与原核表达的P1蛋白特异性结合,ELISA方法检测抗血清的效价可达到1∶5120。结论]该研究结果为建立FMDV的血清学诊断方法及其基因工程疫苗的研究奠定了基础。
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| 关 键 词： | Asia1型口蹄疫病毒 P1基因 原核表达 抗血清 |
Prokaryotic Expression of P1 Gene of Type Asia1 Foot and Mouth Disease Virus (FMDV) and the Preparation of Its Antiserum |
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| WU Gang,WANG Hong-mei,LIU Xiao,WANG Li-qun,YU Li,ZHONG Ji-feng,HE Hong-bin.Prokaryotic Expression of P1 Gene of Type Asia1 Foot and Mouth Disease Virus (FMDV) and the Preparation of Its Antiserum[J].Agricultural Science ＆ Technology,2010(9):112-114,143. |
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| Authors: | WU Gang WANG Hong-mei LIU Xiao WANG Li-qun YU Li ZHONG Ji-feng HE Hong-bin |
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| Institution: | 1.Dairy Cattle Research Center,Shandong Academy of Agricultural Sciences,Jinan 250100;2.College of Life Science,Northeast Agriculture University,Harbin 150030;3.Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150000 |
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| Abstract: | Objective] The aim was to study the prokaryotic expression of P1 gene of foot-and-mouth disease virus（FMDV）type Asia 1and the preparation of its antiserum.Method]The P1 gene of FMDV type Asia 1 was obtained by gene cloning techniques,and then cloned into pET-32a（＋）plasmid;subsequently the recombinant plasmid was transformed into E.coli BL21（DE3）;after the IPTG induction and protein purification,SDS-PAGE analysis was carried out;the ultrasonic wave was use to lyse the cultivated recombinant strain,and after the isolation and purification,this fusion protein was utilized to immunize New Zealand rabbits so as to prepare P1 protein antiserum.Result]The positive clones were obtained;SDS-PAGE result showed that the target band was appeared at 105 kD;Western blot analysis showed that the antisera could bind to the expressed P1 fusion protein specifically;the ELISA titer of the rabbit anti-FMDV-P1 sera was approximately 1∶5 120.Conclusion]This study had provided foundations for FMDV serological diagnostic methods and genetically engineered vaccine. |
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| Keywords: | Foot-and-mouth disease virus（FMDV） P1 gene Prokaryotic expression Antiserum |
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