Ren Xiao-xia, Zhang Xin, Cai Gui-hong, Li Yu-ying, Wang Zhuan-hua
Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2010 Jun;26(6):543-5.
To determine the most important region in tartary buckwheat allergen.
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 Ni(2+);-NTA agarose affinity chromatography column and indirect ELISA, inhibition ELISA and Dot blot was performed using sera from allergenic patients.
The purified proteins were obtained and the immunological results showed that E1 exhibite stronger IgE binding to patient's serum than the other epitopes.
E1 is probably the most important region in tartary buckwheat allergen binding to buckwheat allergic sera IgE.
确定苦荞过敏原中最重要的区域。
根据苦荞过敏蛋白cDNA设计引物，通过PCR扩增表位基因。构建四个含有表位基因的表达载体，然后转化到大肠杆菌BL21(DE3)宿主细胞中。表达产物用Ni(2+)-NTA琼脂糖亲和层析柱纯化，并用过敏患者血清进行间接ELISA、抑制ELISA和斑点印迹。
获得了纯化蛋白，免疫结果显示E1与患者血清的IgE结合比其他表位更强。
E1可能是苦荞过敏原中与苦荞过敏血清IgE结合的最重要区域。
