苦荞过敏原TBa表位区段的表达及免疫活性分析  

Expression and immunological characterization of the epitopes in tartary buckwheat allergen

作　　者：任晓霞[1] 张昕[2] 蔡桂红[1] 李玉英[1] 王转花[1] 

机构地区：[1]山西大学生物技术研究所化学生物学与分子工程教育部重点实验室,山西太原030006 [2]山西大学环境科学与工程研究中心,山西太原030006

出　　处：《细胞与分子免疫学杂志》2010年第6期543-545,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30870525;30671084);山西省自然科学基金资助项目(2007011077)

摘　　要：目的:确定苦荞过敏原(TBa)的主要表位区段。方法:以TBacDNA为模板,利用PCR方法,构建TBa的4个表位区段(E12、E3、E6、E36)的重组原核表达载体,转入大肠杆菌BL21(DE3)中诱导表达。经Ni2+-NTA亲和纯化后,SDS-PAGE分析。采用间接和竞争ELISA以及点杂交技术进行免疫活性测定。结果:获得了纯度较高的TBa重组表位蛋白(E12、E3、E6、E36),与先前获得表位E1、E2的免疫活性比较表明,重组表位蛋白与荞麦过敏症患者血清均有IgE结合活性,其中E1的活性最强。结论:E1可能是苦荞过敏蛋白TBa中最重要的抗原结合区域之一。AIM：To determine the most important region in tartary buckwheat allergen. METHODS： The gene of epitopes was amplified by PCR using the primers designed according to TBa cDNA. Four expression vectors containing the gene of epitopes were constructed, and then transformed into the E.coli BL21（DE3） host cells. The expression products were purified by Ni2＋-NTA agarose affinity chromatography column and indirect ELISA, inhibition ELISA and Dot blot was performed using sera from allergenic patients. RESULTS： The purified proteins were obtained and the immunological results showed that E1 exhibite stronger IgE binding to patient’s serum than the other epitopes. CONCLUSION： E1 is probably the most important region in tartary buckwheat allergen binding to buckwheat allergic sera IgE.

关 键 词：苦荞过敏原 原核表达 ELISA 免疫活性 

分 类 号：Q78[生物学—分子生物学] R392.11[医药卫生—免疫学]