| 猪氨肽酶N多抗血清的制备及活性分析 |
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| 引用本文： | 王衡,刘博奇,任晓峰,宁章勇.猪氨肽酶N多抗血清的制备及活性分析[J].中国兽医科学,2011(8). |
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| 作者姓名： | 王衡 刘博奇 任晓峰 宁章勇 |
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| 作者单位： | 华南农业大学兽医学院;广州中医药大学科技产业园有限公司新南方药物安全性评价中心;东北农业大学动物医学学院; |
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| 摘 要： | 为进一步研究猪氨肽酶N(pAPN)介导TGEV与靶细胞相互作用的机制,将编码猪氨肽酶N成熟肽基因(105～2889bp)克隆至原核表达载体pET-30a,将重组质粒pET-30a-pAPN转化大肠杆菌BL21(DE3),IPTG诱导后进行SDS-PAGE分析,利用蛋白切胶纯化技术对目的蛋白进行纯化,制备其多抗血清,并通过琼扩试验、Western-blot分析、病毒抑制试验及体外瞬时转染试验分析该多抗血清的生物活性。结果,获得了大小为112ku的目的蛋白,多抗血清效价为1∶32,该血清在低浓度时仍具有抑制病毒的活性。结果证实,制备的多抗血清具有较高的抗体效价、较强的特异性及良好的生物活性。
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| 关 键 词： | 猪氨肽酶N(pAPN) 多抗血清 活性检测 |
Biological characteristic analysis of prokaryotic expression of porcine amino peptidase N gene and preparation of its polyclonal antibody |
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| WANG Heng,LIU Bo-qi,REN Xiao-feng,NING Zhang-yong.Biological characteristic analysis of prokaryotic expression of porcine amino peptidase N gene and preparation of its polyclonal antibody[J].Veterinary Science in China,2011(8). |
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| Authors: | WANG Heng LIU Bo-qi REN Xiao-feng NING Zhang-yong |
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| Institution: | WANG Heng1,LIU Bo-qi2,REN Xiao-feng3,NING Zhang-yong1 (1.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China,2.New South Center for Safety Evaluation of Drugs,Science and Technology Park Ltd.of Guangzhou University of TCM,Guangzhou 510540,3.College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | The full-length gene encoding porcine amino peptidase N(pAPN) was cloned into prokaryotic expression vector pET-30a,the recombinant plasmid was identified and designated as pET-30a-pAPN to take further research on the mechanism of the interaction between TGEV and its target cells induced by pAPN. Then pET-30a-pAPN was transformed into E.coli BL21(DE3),and the pAPN was induced expressed by IPTG and analyzed by SDA-PAGE with the protein purified by the method of gel cutting.Agarose diffusion assay,Western-blo... |
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| Keywords: | porcine amino peptidase N(pAPN) antiserum bioactivity detection |
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