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| 德国小蠊Bla g 2的原核表达及其多克隆抗体的制备 | |
| 引用本文： | 张红绪,邢文会,刘丹丹,胡萍,杨萍.德国小蠊Bla g 2的原核表达及其多克隆抗体的制备[J].河南师范大学学报(自然科学版),2010,38(4). |
| 作者姓名： | 张红绪 邢文会 刘丹丹 胡萍 杨萍 |
| 作者单位： | 1. 河南师范大学,生命科学学院,河南,新乡,453007 2. 河南教育学院,人口与生命科学系,郑州,450046 3. 上海我武生物科技有限公司,上海,200233 |
| 基金项目： | 河南省科技攻关项目，河南省动物重点学科资助 |
| 摘 要： | 利用RT-PCR技术扩增德国小蠊Blag2基因,将其克隆进pET-41b载体.在大肠杆菌中经IPTG诱导表达获得GST-Blag2蛋白,SDS-PAGE分析表达产物,得到相对分子量约74kDa的融合蛋白.通过亲和层析法纯化表达的GST-Blag2蛋白,并以此表达产物制成油乳剂疫苗免疫新西兰白兔,制备多克隆抗体.间接ELISA法测得抗体效价达到1∶300000.Western印迹表明抗体有较高的特异性,可分别与GST-Blag2融合蛋白和德国小蠊变应原浸提液反应.Blag2在大肠杆菌中的成功表达及其多克隆抗体的制备,为德国小蠊过敏患者的诊断和治疗提供了帮助. |
| 关 键 词： | Bla g 2 原核表达 多克隆抗体 |
Prokaryotic Expression of Blattella Bermanica Allergen 2(Bla g 2) and Preparation of Polyclonal Antibody |
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| ZHANG Hong-xu,XING Wen-hui,LIU Dan-dan,HU Ping,YANG Ping.Prokaryotic Expression of Blattella Bermanica Allergen 2(Bla g 2) and Preparation of Polyclonal Antibody[J].Journal of Henan Normal University(Natural Science),2010,38(4). | |
| Authors: | ZHANG Hong-xu XING Wen-hui LIU Dan-dan HU Ping YANG Ping |
| Abstract: | Bla g 2 gene is amplified by RT-PCR from the total RNA of Blattella germanica and is inserted into pET-41b vector.GST-Bla g 2 was expressed in E.coli via IPTG induction,a relative molecular mass of approximately 74 kDa is shown by SDS-PAGE.The expressed fusion protein is purified by Ni+-agarose affinity chromatography,an emulsion is prepared with the purified Bla g 2 and injected into rabbits.The titer of antiserum identified by indirect ELISA reached 1∶300 000.Moreover,antiserum reacts specifically with purified fusion protein and the extract of Blattella germanica.Preparation of Bla g 2 and its polyclonal antibody offer help to the dialysis and therapy of patients who are allergic to Blattella germanica. |
| Keywords: | Bla g 2 |
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