华北农学报2017，Vol.32Issue(2)：32-37,6.DOI:10.7668/hbnxb.2017.02.005
掌叶半夏凝集素基因的克隆及亚细胞定位分析
Genomic Cloning an Agglutinin Gene from Pinellia pedatisecta and Analysis on Its Subcellular Localization
摘要
Abstract
To clone Pinellia pedatisecta agglutinin gene(PPA)from the leaves of Pinellia pedatisecta,analyzed on its biological information characteristics,and observed its subcellular localization.The open reading frame sequence of PPA gene was amplified by PCR method.The recombinant vector pCAMBIA1300-35S-PPA-eGFP was constructed and then transferred into Agrobacterium strain GV3101.Agrobacterium suspensions expressing PPA were injected at different concentrations into Nicotiana tabacum leaves and the subcellular location was observed by confocal laser scanning microscope.It showed that the open reading fragment sequence length of PPA gene was 777 bp,encoding 258 amino acids with GenBank accession number KF154981 and AGV40779,respectively.It contained two conservative B-lectin domains and three mannose binding sites.Sequence alignment demonstrated that the deduced amino acid sequence of PPA,which were 96%,91%,86% and 83% identical respectively,with the corresponding region of Arisaema heterophyllum, Pinellia cordata,Arisaema lobatum and Pinellia ternata.The subcellular location results indicated that the PPA-eGFP was located in the plasma membrane as large spots,but not found in nucleus.The results would provide the foundation for further study the structure and function of PPA gene,and elucidate anti-microbial mechanism of PPA.关键词
掌叶半夏/掌叶半夏凝集素/基因克隆/亚细胞定位Key words
Pinellia pedatisecta/Pinellia pedatisecta agglutinin/Gene cloning/Subcellular localization分类
生物科学引用本文复制引用
赵欢..掌叶半夏凝集素基因的克隆及亚细胞定位分析[J].华北农学报,2017,32(2):32-37,6.基金项目
四川省教育厅重大培育项目(15CZ0015) （15CZ0015）
西华师范大学青年教师资助专项(1D012) （1D012）
西华师范大学青年教师资助专项(14D012) （14D012）
