蕙兰TUB基因片段的克隆与序列分析  

Molecular Cloning and Sequence Analysis of TUB Gene Fragments from Cymbidium faberi

作　　者：田云芳[1] 蒋素华[2] 苏金乐[3] 崔波[2] 

机构地区：[1]郑州师范学院生命科学系,郑州450044 [2]郑州师范学院生物研究所,郑州450044 [3]河南农业大学林学院,郑州450002

出　　处：《中国农学通报》2015年第19期117-121,共5页Chinese Agricultural Science Bulletin

基　　金：河南省科技攻关项目"蝴蝶兰的分子育种"(092102110128);河南省教育厅科学技术研究重点项目"蕙兰合适内参基因及开花相关基因的研究"(14A220005)

摘　　要：对蕙兰TUB基因片段进行克隆与序列分析,为蕙兰内参基因的筛选和研究其生理功能提供研究基础。根据NCBI已经公布的TUB基因的同源核苷酸保守序列,进行简并引物设计,利用RT-PCR的方法,以蕙兰花蕾期的花葶为材料,克隆了5条TUB基因片段。序列分析结果表明,5条TUB基因片段长度均为1055 bp,编码351个氨基酸,具有2个TUB基因标记位点:Tubulin-beta m RNA autoregulation signal:MREI和Tubulin subunits alpha,beta,and gamma signature(GGGTGSG)特征信号序列。各片段之间同源性高达91.72%,经BLAST分析,与小麦的TUB1同源性可达85%。利用DNAMAN等生物软件推断的氨基酸,与毛果杨、蓖麻、葡萄同源性达97%。研究中得到的5个基因序列是TUB基因的同源片段,分别命名为Cf TUB1、Cf TUB2、Cf TUB3、Cf TUB4和Cf TUB5,并在Gen Bank注册,登录号分为JN177713、JN177714、JN177715、JN177716和JN177717。TUB of Cymbidium faberi was cloned and analyzed in sequence,and the results would provide animportant basis for selecting a reference gene and studying the function of TUB.According to the conservedsequences of the TUB homologous genes in Gen Bank,a pair of primers was designed.TUB genes wereidentified from the pedicels of C.faberi by homologous cloning.Five TUB genes were successfully cloned andcharacterized.The sequencing result showed that each of the five TUB gene fragments contained 1055 bp,encoding a protein of 351 amino acids.Each TUB contained the signature sequence Tubulin-beta m RNA autoregulation signal（MREI） and Tubulin subunits alpha,beta,and gamma signature（GGGTGSG）.Theyshared 91.72% nucleotide sequence homology with each other and 85% nucleotide sequence homology with Triticum aestivum.The deduced amino acid sequence shared 97% homology with TUB of Populus trichocarpa,Ricinus communis,Vitis vinifera.The cloned genes were TUB gene fragments,named as Cf TUB1,Cf TUB2,Cf TUB3,Cf TUB4 and Cf TUB5,respectively.They were registered into Gen Bank（respective accession number：JN177713,JN177714,JN177715,JN177716 and JN177717）.

关 键 词：蕙兰 TUB 克隆 序列分析 

分 类 号：Q781[生物学—分子生物学]