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颗粒结合型淀粉合成酶Ⅰ(granule-bound starch synthaseⅠ,GBSSⅠ)是决定果实直链淀粉合成的关键酶。研究GBSSⅠ的亚细胞定位及其在毕赤酵母中的表达,将为其蛋白功能验证奠定基础。本研究从巴西蕉(Musa acuminata L.AAA group cv.Brazilian)果实中克隆到一个GBSSⅠ成员,命名为Ma GBSSⅠ-3。生物信息学分析表明,Ma GBSSⅠ-3开放阅读框为675 bp,编码224个氨基酸,蛋白分子量为55.12 k D,等电点为5.38。聚类分析发现Ma GBSSⅠ-3与油棕Eg GBBSⅠ的亲缘关系较近。亚细胞定位显示,Ma GBSSⅠ-3定位于细胞膜。酵母表达系统分析发现,Ma GBSSⅠ-3蛋白的大小约为55.0 k D,与预测的分子量大小相一致,说明已成功获得了Ma GBSSⅠ-3表达蛋白,为进一步验证Ma GBSSⅠ-3蛋白的功能奠定了基础。
Granule-bound starch synthaseⅠ (GBSSⅠ) is the key enzyme that determines amylose synthesis in fruit. Studying the subcellular location of GBSSⅠ and its expression in Pichia pastoris will lay the foundation for its protein functional verification. In this study, a member of GBSSⅠ was cloned from the fruit of Musa acuminata L.AAA group cv.Brazilian and named Ma GBSSⅠ-3. Bioinformatics analysis showed that the open reading frame of Ma GBSSⅠ-3 was 675 bp, encoding 224 amino acids with a molecular weight of 55.12 kD and an isoelectric point of 5.38. Cluster analysis showed that Ma GBSSⅠ-3 was more closely related to EgGB EgⅠ. Subcellular localization showed that Ma GBSS I-3 localized to the cell membrane. Yeast expression system analysis showed that the size of Ma GBSSⅠ-3 protein was about 55.0 kD, which was consistent with the predicted molecular weight, indicating that Ma GBSSⅠ-3 protein was successfully obtained, which laid the foundation for further verifying the function of Ma GBSSⅠ-3 protein The foundation.