华北农学报2017，Vol.32Issue(3)：58-64,7.DOI:10.7668/hbnxb.2017.03.009
芍药分生组织决定基因APETALA2(AP2)的克隆及生物信息学分析
Cloning and Bioinformatics Analysis of APETALA2 (AP2) in Paeonia lactiflora
摘要
Abstract
Flower-meristem-identity gene APETALA2(AP2) belonging to ABCDE model plays a role in regulating the development of plant floral organ.In order to understand the biological function of P.lactiflora AP2 gene,the plAP2 gene sequence of Paeonia lactiflora was cloned using RACE and sequencing technique technology,meanwhile the sequence signature,protein structure and function,subcellular localization were analyzed by bioinformatics methods.Molecular phylogenetic tree of AP2 from different plants were constructed by MEGA 5.0 software.Finally,we detected the differential expression between inner-petal and outer-petal by qPCR.Results showed,the full-length cDNA sequence of plAP2 gene was 1 935 bp and consisted of a 1 578 bp open reading frame (ORF) encoding 525 amino acid proteins.Analysis of protein structure and function showed,plAP2 was a hydrophilic and unstable protein,meanwhile plAP2 was also non-secretory protein without signal peptide and transmembrane structure.Nuclear localization signal located between 139th and 147th amino acid (KKSRRGPRS).The secondary structure of plAP2 protein contained 24% α-helices,19% β-sheet,28% β-turn and 28% random coil.plAP2 protein had eight glycosylation sites and sixty-four phosphorylation sites.plAP2 protein had two same conservative domain:AP2/ Ethylene-Responsive factors (ERF), which was located in the region between 151 and 213,between 243 and 306,respectively.Prediction of subcellular localization showed that plAP2 located mostly in cytoplasm (45.0%),rarely in microbody, mitochondrial matrix and lysosome.Analysis of evolutionary tree showed that the AP2 of Paeonia lactiflora were highly homologous and close to Paeonia suffruticosa.qPCR showed the expression of AP2 in outer-petal was extremely significant than that in inner-petal (P<0.01).In this study,we cloned the complete cDNA sequence of AP2 gene in Paeonia lactiflora and systematically revealed the basic protein structure,functional sites,domains,cellular localization and expression level,which will provide basic materials and theoretical references for further studying plAP2 gene function.关键词
芍药/APETALA2基因/RACE/生物信息学Key words
Paeonia lactiflora/APETALA2 gene/RACE/Bioinformatics分类
生物科学引用本文复制引用
吴彦庆,成梦琳,赵大球,陶俊..芍药分生组织决定基因APETALA2(AP2)的克隆及生物信息学分析[J].华北农学报,2017,32(3):58-64,7.基金项目
国家自然科学基金项目(31372097 （）
31400592) （）
江苏省农业科技自主创新资金项目(CX(12)2019) （CX(12）
