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以成熟的黄果柑果实为材料,根据已登录植物的XET基因并参考甜橙基因组设计特异性引物,采用同源克隆方法,获得黄果柑XET基因完整的编码区序列,命名为Cit XET。运用生物学软件对该序列及其编码的氨基酸序列进行相关生物信息学及结构功能分析,结果显示黄果柑XET基因长960 bp,与其它植物序列具有较高的同源性。该基因编码319个氨基酸,蛋白质分子量为37.4547 k D,等电点为9.05,分子式为C_(1724)H_(2548)N_*(448)O_(466)S_(14),是一个具有跨膜结构的亲水性蛋白。序列比对及进化树分析,XET在进化的过程中具有高度的保守性。同时,Cit XET蛋白二级结构有21%的α-螺旋,30.72%的延伸链,9.09%的β-转角,39.18%的无规蜷曲。黄果柑XET具有GH16等结构域,属于糖苷水解酶GH16超家族的成员,其含有该家族的特征基序DEIDFEFLG和β-卷芯(β-jellyroll)折叠结构,同时预测该蛋白质可能参与碳水化合物代谢,木葡聚糖代谢,细胞壁生物合成等过程。本研究分离鉴定了黄果柑XET基因,同时初步了解该基因的生物学信息,有助于进一步探讨其的表达模式和具体功能,为黄果柑果实品质的改善提供理论依据。
Based on the XET gene of the registered plant and the specific primers designed by referring to the sweet orange genome, the full-length coding sequence of XET gene of Citrus grandis was obtained by homologous cloning and named Cit XET. Bioinformatics and structural function analysis of this sequence and its encoded amino acid sequence using biological software showed that the XET gene of Huangguogurt was 960 bp long and shared high homology with other plant sequences. The gene encodes a protein of 319 amino acids with a molecular mass of 37.4547 kD, an isoelectric point of 9.05 and a molecular formula of C_ (1724) H_ (2548) N_ * (448) O_ (466) S_ (14) Hydrophilic protein. Sequence alignment and phylogenetic tree analysis, XET in the evolutionary process is highly conserved. At the same time, Cit XET protein secondary structure has 21% alpha-helix, 30.72% extension chain, 9.09% beta-turn and 39.18% random twist. XET has the domain of GH16 and belongs to the GH16 superfamily of glycoside hydrolase, which contains the characteristic motif DEIDFEFLG and β-jellyroll folding structure of this family, and predicts that the protein may be involved in carbohydrate Metabolism, xyloglucan metabolism, cell wall biosynthesis and other processes. In this study, XET gene was isolated and identified, and its biological information was preliminarily understood. It is helpful to further explore the expression pattern and function of this gene and provide a theoretical basis for the improvement of fruit quality.