小麦γ-醇溶蛋白TaWG05基因克隆、原核表达及多克隆抗体制备

Clonging, Prokaryotic Expression and Polyclonal Antibody Preparation of γ-Gliadin Gene TaWG05 in Triticum aestivum

醇溶蛋白是小麦储藏蛋白的重要组成成分,也是一种主要的食物过敏原,可引发乳糜泻和食物依赖运动激发等多种症状。本研究以郑麦004为材料,克隆得到一个γ-醇溶蛋白Ta WG05基因,该基因编码319个氨基酸,其33~40氨基酸残基位置存在Ig E抗原表位区段,可能与小麦依赖运动激发过敏症有关。制备Ta WG05蛋白的抗体,利用Western Blot技术在不同小麦品种中检测Ta WG05蛋白的表达情况,发现在郑麦366、郑麦9023和新麦26中,Ta WG05蛋白几乎无表达;而在郑麦004、矮抗58、豫麦18和偃展4110中,其表达量较高。所得数据表明该抗体可以有效的识别Ta WG05蛋白在小麦品种中的表达情况,为该蛋白过敏人群提供较好的健康指导,也为相关品种改良提供理论参考和技术支撑。

Gliadin is a main component of wheat storage proteins, and it is also a major food allergen, which could cause many symptoms such as celiac disease and wheat-dependent exercise-induced anaphylaxis. In this study, we cloned a γ-gliadin gene TaWG05 from Zhengmai004, which encoded 319 amino acids. Bioinformatics analysis found that the 33~40 amino acid residues contained an Ig E-binding epitope, and it might be related with wheat-dependent exercise-induced anaphylaxis. Furthermore, we prepared the antibody of TaWG05（α-TaWG05）and used Western Blot to detect the expression of TaWG05 in different varieties of wheat. The results showed that the expression of TaWG05 was not detected in Zhengmai 366, Zhengmai 9023 and Xinmai26, but it had high expression in Zhengmai 004, Aikang 58, Yumai 18 and Yanzhan 4110. In conclusion, α-TaWG05 could effectively identify TaWG05 protein expression in wheat varieties, which could provide good health guidance for allergy crowd of this protein and afford theoretical basis and technical support for improvement of related wheat varieties.