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中国农学通报

中国农学通报 ›› 2015, Vol. 31 ›› Issue (19): 117-121.doi: 10.11924/j.issn.1000-6850.2014-1356

所属专题： 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

蕙兰TUB基因片段的克隆与序列分析

田云芳,蒋素华,苏金乐,崔波   

  1. 郑州师范学院,郑州师范学院,河南农业大学,郑州师范学院
  • 收稿日期:2014-05-12 修回日期:2015-06-18 接受日期:2014-08-26 出版日期:2015-07-28 发布日期:2015-07-28
  • 通讯作者: 田云芳
  • 基金资助:
    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)。

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

  • Received:2014-05-12 Revised:2015-06-18 Accepted:2014-08-26 Online:2015-07-28 Published:2015-07-28

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1055bp，编码351个氨基酸，具有2个TUB基因标记位点：Tubulin-betamRNAautoregulationsignal：MREI和Tubulinsubunitsalpha,beta,andgammasignature(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CfTUB1、CfTUB2、CfTUB3、CfTUB4和CfTUB5，并在GenBank注册，登录号分为JN177713、JN177714、JN177715、JN177716和JN177717。

关键词: 发展, 发展

Abstract: TUB of Cymbidium faberi was cloned and analyzed in sequence, and the results would provide an important basis for selecting a reference gene and studying the function of TUB. According to the conserved sequences of the TUB homologous genes in GenBank, a pair of primers was designed. TUB genes were identified from the pedicels of C. faberi by homologous cloning. Five TUB genes were successfully cloned and characterized. 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 mRNA autoregulation signal (MREI) and Tubulin subunits alpha, beta, and gamma signature (GGGTGSG). They shared 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 CfTUB1, CfTUB2, CfTUB3, CfTUB4 and CfTUB5, respectively. They were registered into GenBank(respective accession number: JN177713, JN177714, JN177715, JN177716 and JN177717).