掌叶半夏凝集素基因的克隆及亚细胞定位分析

发布时间：2018-06-04 05:07

本文选题：掌叶半夏 + 掌叶半夏凝集素　；参考：《华北农学报》2017年02期

【摘要】：为了克隆掌叶半夏凝集素基因(Pinellia pedatisecta agglutinin,PPA),并对其进行生物信息学分析和亚细胞定位。采用PCR技术扩增掌叶半夏凝集素基因的ORF序列,构建pCAMBIA1300-35S-PPA-e GFP重组载体,导入农杆菌GV3101并在烟草中进行瞬时表达,采用激光扫描共聚焦显微镜观察其亚细胞定位。克隆获得的PPA基因全长由777个核苷酸组成,编码258个氨基酸,含有3个甘露糖结合位点和2个保守的B-lectin结构域,Gen Bank登录号为KF154981,氨基酸序列登录号为AGV40779。序列比对表明,该序列与天南星、滴水珠、花南星和半夏凝集素的氨基酸序列相似性分别为96%,91%,86%和83%。亚细胞定位的结果表明,pCAMBIA1300-35S-PPA-e GFP融合蛋白在细胞核内无分布,而是点状分布在细胞质膜上。为进一步解析掌叶半夏凝集素基因的结构与功能,开展掌叶半夏凝集素抗病虫害机制的研究奠定基础。
[Abstract]:In order to clone Pinellia pedatisecta agglutininine (PPAA), the lectin gene of Pinellia ternata was analyzed by bioinformatics and subcellular localization. The ORF sequence of the lectin gene of Pinellia ternata was amplified by PCR, and the recombinant vector of pCAMBIA1300-35S-PPA-e GFP was constructed. The recombinant vector was introduced into Agrobacterium tumefaciens GV3101 and expressed instantaneously in tobacco. The subcellular localization was observed by laser scanning confocal microscopy. The cloned PPA gene is composed of 777 nucleotides and encodes 258 amino acids. It contains three mannose binding sites and two conserved B-lectin domain Genin Bank accession number is KF154981, and the amino acid sequence accession number is AGV40779. The sequence alignment showed that the amino acid sequence similarity between the sequence and the Amino acid sequence of Tiananxing, droplets, Pternata and Pinellia ternata were 961.86% and 833%, respectively. The results of subcellular localization showed that pCAMBIA1300-35S-PPA-e GFP fusion protein was not distributed in the nucleus, but in the cytoplasmic membrane. In order to further analyze the structure and function of the lectin gene of Pinellia ternata and to develop the mechanism of resistance to diseases and insect pests of Pinellia ternata lectin.
【作者单位】：西华师范大学生命科学学院;
【基金】：四川省教育厅重大培育项目(15CZ0015) 西华师范大学青年教师资助专项(1D012);西华师范大学青年教师资助专项(14D012)
【分类号】：S567.239

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