| 恶性疟原虫肝素结合蛋白质PF3D7_1104400的原核表达与多克隆抗体的制备 |
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| 引用本文： | 黄朋,赵帅杰,陆慧君,尹继刚,姜宁,陈启军.恶性疟原虫肝素结合蛋白质PF3D7_1104400的原核表达与多克隆抗体的制备[J].吉林农业大学学报,2017,39(1). |
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| 作者姓名： | 黄朋 赵帅杰 陆慧君 尹继刚 姜宁 陈启军 |
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| 作者单位： | 吉林大学人兽共患病研究所,人兽共患病研究教育部重点实验室,长春130062 |
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| 摘 要： | 为研究肝素结合蛋白质PF3D7_1104400的功能以及其作为疟疾疫苗候选抗原的可行性,通过PCR方法扩增目的基因,并将其与p ET-28a和p GEX-4T-1表达载体连接构建重组表达质粒。将构建成功的重组表达质粒转化入大肠杆菌BL21-Condon Plus(DE3)-RIPL中诱导表达,利用亲和层析的方法纯化His-tag和GSTtag重组蛋白质。用His-tag重组蛋白质免疫家兔制备多克隆抗体,以该抗体为一抗,利用Western blot检测该抗体的特异性以及PF3D7_1104400蛋白质在虫体内是否表达。结果表明:成功构建重组表达质粒PF3D7_1104400-p ET和PF3D7_1104400-p GEX,诱导表达并纯化出特异性好、纯度高的重组蛋白。制备的多克隆抗体能够特异性识别虫体天然蛋白,PF3D7_1104400蛋白质在恶性疟原虫体内全长表达。
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| 关 键 词： | 恶性疟原虫 肝素 原核表达 多克隆抗体 疫苗候选抗原 |
Prokaryotic Expression and Preparation of Polyclonal Antibody of Heparin Binding Protein PF3D7_1104400 of Plasmodium falciparum |
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| HUANG Peng,ZHAO Shuaijie,LU Huijun,YIN Jigang,JIANG Ning,CHEN Qijun.Prokaryotic Expression and Preparation of Polyclonal Antibody of Heparin Binding Protein PF3D7_1104400 of Plasmodium falciparum[J].Journal of Jilin Agricultural University,2017,39(1). |
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| Authors: | HUANG Peng ZHAO Shuaijie LU Huijun YIN Jigang JIANG Ning CHEN Qijun |
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| Abstract: | To research the biological function of heparin binding protein PF3D7_1104400 and its feasibility as candidate vaccine antigen,target gene was amplified by PCR whose products were con nected with pET-28a and pGEX-4T-1 for constructing recombinant expression plasmids.Transformed into BL21-CondonPlus(DE3)-RIPL,recombinant plasmids were induced and expressed and recombinant proteins were purified by the method of affinity chromatography.Polyclonal antibody was prepared by rabbits immuned with His-tagged recombinant proteins.The specificity of antibody and expression of PF3D7_1104400 were detected by Western blot in which polyclonal antibody was used as primary antibody.The results showed that recombinant plasmids,PF3D7_1104400-pET and PF3D7_1104400-pGEX,were constructed successfully and specific recombinant proteins were expressed and purified.According to the results of Western blot,natural protein of PF3D7_1104400 could be recognized specifically by prepared antibody and was expressed in Plasmodium falciparum in the form of total length,which will lay foundation for further research of PF3D7_1104400. |
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| Keywords: | plasmodium falciparum heparin prokaryotic expression polyclonal antibody candidate vaccine antigen |
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