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植物的同源异型盒基因(knotted1-like homeobox,KNOX)与叶片形态建成具有密切的相关性。本研究以剑麻(Agave sisalana)茎顶端分生组织为材料分离得到编码357个氨基酸的As KNOX I全长基因,预测该蛋白的分子量约为40 k Da,等电点为p I 6.4,且包含KNOX I同源蛋白家族典型的HD、MEINOX、ELK和GSE保守结构域。后通过构建原核和真核表达载体,分别在大肠杆菌和烟草叶片中瞬时表达,用His标签进行蛋白纯化,Western blot用于表达产物检测。结果表明,As KNOX I基因已在原核和真核中成功表达,为进一步分析该蛋白的结构及功能特征提供了技术支持。
Plant homeobox gene (Knotted1-like homeobox, KNOX) and leaf morphogenesis is closely related. In this study, the full-length As KNOX I gene encoding 357 amino acids was isolated from the apical meristem of Agave sisalana. The predicted molecular weight of As KNOX I was about 40 kDa and the isoelectric point was p I 6.4. Contains the conserved domains of HD, MEINOX, ELK and GSE typical of the KNOX I homologue family. After construction, prokaryotic and eukaryotic expression vectors were constructed and transiently expressed in Escherichia coli and tobacco leaves respectively. His tag was used for protein purification. Western blot was used to detect the expression product. The results showed that As KNOX I gene was successfully expressed in prokaryotic and eukaryotic cells, providing technical support for further analysis of the structural and functional characteristics of the protein.