大米蜡质蛋白的原核表达、抗体制备及检测

Prokaryotic Expression of Rice Waxy Protein,Antibody Preparation and Detection

大米蜡质蛋白是大米胚乳中直链淀粉合成的关键酶。它是大米的主要致敏原之一。为了建立起大米蜡质蛋白的检测方法,本研究对大米蜡质蛋白的编码基因进行了亚克隆,并用大肠杆菌合成了蜡质蛋白的重组蛋白片段。通过已纯化的重组蜡质蛋白片段为抗原免疫白兔,制备了多克隆抗体。该抗体能够特异性地与大米蜡质蛋白进行免疫结合反应,并且,利用该抗体对大米蜡质蛋白质进行了Western-blotting检测和ELISA检测。

Rice waxy protein is a key enzyme in synthese of amylose in the endosperm of rice grain, which is also one of the main allergens in rice. In order to develop methods to detect this protein, the gene of rice waxy protein was sub-cloned and expressed in E.coli, and antibodies were prepared by immunizing rabbits with recombinant rice waxy protein fragment. The antibodies specifically immune reacted with rice waxy protein. Moreover, the antibody was applied to detect and analysis rice waxy protein with Western-blotting and ELISA.