【摘要】 为进一步解析γ-醇溶蛋白基因在小麦面粉品质中的调控机制，本研究从郑麦158(低蛋白质含量、高面团强度)中克隆了小麦γ-醇溶蛋白基因Tagli-γ-11。生物信息学分析结果表明，该基因具有γ-醇溶蛋白的典型结构特征，含有8个保守的半胱氨酸残基，并在该蛋白质的重复区Ⅱ内第133～141 aa位置发现1个乳糜泻(CD)表位(PQQSFPQQQ)。利用酵母双杂交技术筛选郑麦158的cDNA文库，共筛选了8个可能与Tagli-γ-11互作的蛋白质，它们分别是半胱氨酸蛋白酶、果糖二磷酸醛缩酶、富含半胱氨酸和跨膜结构域蛋白1、(1,3∶1,4)-β-D-葡聚糖酶、生长素响应因子ARF17-like、细胞数目调控因子CNR8-like、泛素结构域蛋白DSK2b和转录因子PIF1-like。3个代表性候选蛋白质[富含半胱氨酸和跨膜结构域蛋白1、果糖二磷酸醛缩酶、(1,3∶1,4)-β-D-葡聚糖酶]的回转验证结果表明，它们与Tagli-γ-11均存在互作关系，推测Tagli-γ-11与这些蛋白质相互作用，主要参与了小麦籽粒中贮藏蛋白(如醇溶蛋白)和淀粉的合成与降解，以及生殖生长过程。更多还原

【Abstract】 To further analyze the regulation mechanism of γ-gliadin gene in flour quality of wheat, the Tagli-γ-11 gene was successfully cloned from Zhengmai 158(low protein content and high dough strength). Bioinformatic analysis revealed that this gene possessed the typical structural characteristics of γ-gliadin, containing eight conserved cysteine residues. A celiac disease(CD) epitope(PQQSFPQQQ) was identified within repeat domain Ⅱ of the Tagli-γ-11 protein, located at amino acid positions 133-141. Using the yeast two-hybrid(Y2H) system, the cDNA library of Zhengmai 158 was screened, yielding eight candidate proteins potentially interacting with Tagli-γ-11. These included cysteine protease, fructose-bisphosphate aldolase, cysteine-rich and transmembrane domain-containing protein 1,(1,3∶1,4)-β-D-glucanase, auxin response factor 17-like(ARF17-like), cell number regulator 8-like(CNR8-like), ubiquitin-associated domain-containing protein DSK2b, transcription factor PIF1-like. The rotation validation assays performed on cysteine-rich and transmembrane domain-containing protein 1, fructose-bisphosphate aldolase, and(1,3∶1,4)-β-D-glucanase confirmed physical interaction with Tagli-γ-11. Based on these results, it was hypothesized that Tagli-γ-11 interacted with these proteins and participated in the synthesis and degradation of storage proteins(such as gliadin) and starch within wheat grains, as well as in reproductive growth processes.更多还原