华北农学报2018，Vol.33Issue(1)：109-114,6.DOI:10.7668/hbnxb.2018.01.017
掌叶半夏凝集素基因克隆及原核表达
The Agglutinin Gene Cloning of Pinellia pedatisecta and Prokaryotic Expression
摘要
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.Bioinformat-ics 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.Con-structed 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.关键词
掌叶半夏/凝集素/载体构建/原核表达Key words
Pinellia pedatisecta/Lectin/Vector construction/Prokaryotic expression分类
生物科学引用本文复制引用
王洪乐,齐连芬,杨超沙,吴志明,李亚栋..掌叶半夏凝集素基因克隆及原核表达[J].华北农学报,2018,33(1):109-114,6.基金项目
河北省应用基础研究计划重点基础研究项目(14966503D) （14966503D）
河北省农林科学院博士基金项目(F15R01) （F15R01）
