华北农学报2012，Vol.27Issue(5)：1-7,7.
枳2个F-box基因cDNA全长的克隆及其亚细胞定位分析
Cloning and Subcellular Localization Analysis of Two F-box Genes from Poncirus trifoliata
摘要
Abstract
A bioinformatics strategy is applied to clone a full length cDNA of TIR1 and Pt-F-box gene from citurs by blasting search of EST database with homologous gene cDNA of Arabidopsis thaliana and identified. According to the cDNA of above sequence,the 5'-end and 3'-end sequence were obtained from cDNA library of opening flower of Poncirus trifoliata (L. ) Raf. Using two gene specific primers by 5'RACE and 3'RACE methods,respectively. The full length cDNA of TIR1 and F-box gene from Poncirus trifoliata was spliced based on 5'-end and 3'-end sequence. The complete cDNA,designated as Pt-TIR1 and Pt-F-box,were 2 048,1 695 bp,respectively. These sequences were deposited in GenBank database,accession numbers FJ502240 and FJ502241 ,with an open reading frame encoding 569 and 468 amino acids,respectively. T Bioinformatics analysis showed that the cDNA of Pt-TIRl and Pt-F-box have the recognition sites of microRNA393 and microRNA394. Pt-TIRl and Pt-F-box with other plant F-boxes have the same ami-no acid sequences that are highly conserved as the designed F-box domains. Recombinant plasmid 35S-GW-FJ502240/FJ502241-GFP was introduced into onion epidermal cells by the particle bombardment method with a PDS1000/He. Transformed cells were incubated for 24 h at 221 in the dark and green fluorescence was monitored under a laser scanning confocal microscope. Subcellular localization results showed that the Pt-TIRl and Pt-F-box were localized in the nucleus.关键词
枳/TIR1和F-box/基因克隆/亚细胞定位Key words
Poncirus irifoliata/TlR1 and F-box/ Cloning /Subcellular localization分类
农业科技引用本文复制引用
张演义,王化坤,宋长年,孙欣,王西成..枳2个F-box基因cDNA全长的克隆及其亚细胞定位分析[J].华北农学报,2012,27(5):1-7,7.基金项目
教育部科学技术研究重点项目(109084) （109084）
江苏省2009年度高校研究生科研创新计划(CX09B_238Z) （CX09B_238Z）
