龙眼胚性愈伤组织微管蛋白基因β-tubulin克隆及序列分析  

Cloning and sequence analysis of β-tubulin gene from embryogenic callus in Dimocarpus longan Lour.

作　　者：王亚婷[1] 林玉玲[1] 曾丽兰[1] 赖钟雄[1] 

机构地区：[1]福建农林大学园艺植物生物工程研究所,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2014年第1期49-54,共6页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金资助项目(31078717;31279142);福建省重大科技平台建设资助项目(2008N2001)

摘　　要：以龙眼松散型胚性愈伤组织为材料,利用RT-PCR法首次克隆2条含有完整开放阅读框的β-tubulin基因的cDNA序列,命名为dl-β-tubulin3和dl-β-tubulin6,长度分别为1350和1351 bp,均编码含有446个氨基酸残基的蛋白质.生物信息学分析表明,该蛋白属于亲水性蛋白,无跨膜结构域;与毛果杨、大豆、水稻、玉米和拟南芥具有较高的同源性;含有微管蛋白所特有的2个保守结构域NNWAKGH和RKAFLHWYTGEGMDEMEFTE,以及1个GTP结合位点GGGTGSG.推测β-tubulin通过不同的磷酸化方式改变酶活性及蛋白构象,参与龙眼体胚发生过程的调控.Embryogenic callus was used as the material for cloningβ^-tubulin gene by RT-PCR method in Dimocarpus longan Lout. By the RT-PCR method, two β-tubulin gene sequence were firstly cloned from Dimocarpus longan Lout, which were named dl-β-tu- bulin3 and dl-β-tubulin6, respectively. Their length were 1350 and 1351 bp, which all included a whole open reading frame （ORF） encoding a protein of 446 amino acid residues. Bioinformatic analysis showed that the β-tubulin protein all were hydrophilic, without the signal peptide and transmembrane helices; and they had high amino acid sequence homology with the tubulins in other species such as Populus trichocarpa, Glycine max, Ory＇za sativa, Zea mays and Arabidopsis thaliana ; besides the predicted β-tubulin protein contained two conserved amino acid sequence motifs NNWAKGH, RKAFLHWYTGEGMDEMEFTE and a GrIp binding site GGGTGSG. It was inferred that the β-tubulin was involod in the process of somatic embryogenesis by the various phesphorylation patterns for changing the emzyme activity and protein structure.

关 键 词：龙眼 胚性愈伤组织 微管蛋白基因 克隆 序列分析 

分 类 号：S667.2[农业科学—果树学]