番茄Dof转录因子SlDof1的克隆及表达分析.doc

文档介绍：
番茄 Dof 转录因子 SlDof1 的克隆及表达
分析#
张建苓,陈,杜利欣,朱明库,董婷婷,胡宗利**
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(重庆大学生物工程学院,重庆 400044)
摘要:根据已经报道的拟南芥、玉米中的 Dof 蛋白,通过同源比对,在 NCBI 数据库中筛选
得到了一个 Dof 同源蛋白, 命名为 SlDof1 ( XP_004233366 ), 并根据其基因序列
(XM_004233318)设计特异引物,用 RT-PCR 方法从野生型(WT)番茄(AC++)中克隆得到
了该基因的 cDNA 全长序列。利用生物信息学和实时定量 PCR(RT-PCR)方法,对 SlDof1
进行了生物信息学分析、表达模式分析、胁迫及激素处理表达分析,结果表明:1)SlDof1
基因 CDS 全长 1802 bp,其中包括 1404 bp 的开放阅读框(ORF),编码 467 个氨基酸残基;2)
实时定量 PCR(RT-PCR)表达模式分析结果表明 SlDof1 可能在 WT 番茄叶片发育及果实
成熟过程中发挥着重要的调控作用;3)胁迫及激素处理的 RT-PCR 表达分析结果表明:在
机械损伤处理和高盐处理 WT 番茄的叶子及根时,该基因均受到不同程度的诱导表达,且其
表达明显受外源激素 ABA、ACC、IAA、GA3 和 ZT 的影响。这些结果为进一步研究 SlDof1
在番茄生长发育过程中所起到的重要生理功能奠定了基础。
关键词:番茄;Dof 转录因子;SlDof1;激素及胁迫处理;RT-PCR 分析
中图分类号: Q78;
Cloning and expression analysis of a Dof transcription factor,
SlDof1 in Solanum lycopersicum
ZHANG Jianling, CHEN Guoping, DU Lixin, ZHU Mingku, DONG Tingting,
HU Zongli
(College of Bioengineering, Chongqing University, ChongQing 400044)
Abstract: In this paper,a Dof homologous protein was selected from the NCBI Database by means
of program of homologous alignment according to the published Dof protein in Arabidopsis
thaliana and Maize and named SlDof1(XP_004233366), and then the specific primers, being used
to amplify the cDNA sequences of SlDof1 (XM_004233318)from wild type(WT) tomato(AC++),
were designed according to the DNA corresponding sequences. Bioinformatics analysis ,
expression pattern analysis and analysis of stress and hormone treatment were used to study the
Dof gene SlDof1 utilizing the methods of bioinformatics and Real-time PCR. The results of
bioinformatics analysis indicated that the full length cDNA of SlDof1 was 1802 bp, whose open
reading frame was 1404 bp, encoding 467 amino acid residue. The Real-time PCR (RT-PCR)
expression pattern analysis showed that SlDof1 may play important roles in leaves development
and fruit ripening in WT tomato. The RT-PCR results of stress and hormone treatment showed
that the expression of S