摘要： 为了获得新的掌叶半夏凝集素家族基因，以掌叶半夏叶片为材料，根据GenBank中已报道的天南星科凝集素基因序列设计引物，PCR法扩增获得3个掌叶半夏凝集素基因，分别暂命名为PPA15、PPA324、PPA533。其中，PPA15的全长为729 bp，编码243个氨基酸；PPA324的全长为774 bp，编码258个氨基酸；PPA533的全长为777 bp，编码259个氨基酸。利用分析软件和网站等工具对克隆的3个基因进行生物信息学分析，结果显示，PPAs与GenBank中收录的天南星科半夏基因具有较高同源性，同时具有单子叶植物特有的甘露糖结合位点，推测它们属于同一基因家族。克隆的3个基因均具有信号肽特征，由N端25个氨基酸残基组成，具有典型的跨膜结构域，推测定位于胞内膜结构。构建pET-28b （+）-PPAs原核表达载体，利用大肠杆菌BL21（DE3）进行原核诱导表达，SDS-PAGE结果表明，重组蛋白分子量约28.0 ku，与预期一致。试验结果为进一步研究掌叶半夏凝集素家族蛋白的功能奠定了基础。

关键词: 掌叶半夏, 凝集素, 载体构建, 原核表达

Abstract: To obtain the novel menbers of Pinellia pedatisecta lectin gene family,according to Arisaema lectin gene sequence reported in GenBank,three agglutinin genes were amplified by PCR with designed primers from DNA extracted from leaves of Pinellia pedatisecta,temporarily named PPA15, PPA324, PPA533.The full length of PPA15, PPA324 and PPA533 were 729,774,777 bp,respectively,encoded 243,258,259 amino acids.Bioinformatics analysis showed that the PPAs were high homology with Araceae pinellia genes of GenBank,moreover,they all had mannose unique binding sites in monocotyledonous plants,suggesting that they might belong to the same gene family.The genes were characterized by signal peptide,which consisted of 25 amino acid residues at the N terminal.They had typical transmembrane domain and were presumed to be localized in the inner membrane structure.Constructed the prokaryotic expression vectors pET-28b (+)-PPAs,the recombinant proteins expressed in Escherichia coli BL21 (DE3),moreover,results of SDS-PAGE showed that the recombinant protein had molecular weight about 28.0 ku,which was consistent with the expectation.The results laid the foundation for the further study of Pinellia pedatisecta lectin family proteins function.

Key words: Pinellia pedatisecta, Lectin, Vector construction, Prokaryotic expression

中图分类号: 

• Q78

• S567.03