[1]何慧,丁一琼,汪潇琳,等.野生大豆GsADF1基因的克隆与表达分析[J].大豆科学,2009,28(05):784-790.[doi:10.11861/j.issn.1000-9841.2009.05.0784]
　HE Hui,DING Yi-qiong,WANG Xiao-lin,et al.Cloning of the GsAF1 Gene and Expression Analysis From Soybean (Glycine soja)[J].Soybean Science,2009,28(05):784-790.[doi:10.11861/j.issn.1000-9841.2009.05.0784]
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野生大豆GsADF1基因的克隆与表达分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第28卷 期数: 2009年05期 页码: 784-790 栏目: 出版日期: 2009-10-25
Title:
Cloning of the GsAF1 Gene and Expression Analysis From Soybean (Glycine soja)
文章编号:
1000-9841(2009)05-0784-07
作者:
何慧; 丁一琼; 汪潇琳; 喻德跃
南京农业大学 大豆研究所，国家大豆改良中心，作物遗传与种质创新国家重点实验室，江苏 南京 210095
Author(s):
HE Hui; DING Yi-qiong; WANG Xiao-lin; YU De-yue
Soybean Research Institute，Nanjing Agricultural University，National Center for Soybean Improvement，National Key Laboratory of Crop Genetics and Germplasm Enhancement，Nanjing 210095，Jiangsu，China
关键词:
野生大豆; 肌动蛋白解聚因子; GsADF1; 克隆
Keywords:
Soybean (Glycine soja); Actin depolymerizing factor; GsADF1; Cloning
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2009.05.0784
文献标志码:
A
摘要:
ADF蛋白是一种在真核生物中广泛存在的低分子量的肌动蛋白结合蛋白，在细胞分裂、细胞运动、植物顶端生长如花粉管伸长，根毛的形成等重要的生命活动中发挥着重要的作用。以野生大豆(Glycine soja)的叶片cDNA为模板，通过RT-PCR方法扩增分离获得GsADF1基因。该基因全长为 693 bp，包含417 bp的开放阅读框;其编码的ADF蛋白含139个氨基酸残基。运用生物信息学方法将GsADF1与其他物种ADF蛋白氨基酸序列进行多重比对并构建系统进化树，发现GsADF1与其它生物的ADF蛋白具有很高的同源性。这说明植物ADF基因高度保守。为进一步研究GsADF1在野生大豆不同组织，器官以及不同发育期大豆种子中的表达情况，对它进行了荧光定量分析(Real time RT-PCR)。结果表明：GsADF1在野生材料的根、茎、叶、花和种子中都有表达，且以根和花中表达量最高。而对其在不同发育阶段的种子中的分析表明该基因在15DAF、20DAF、30DAF、40DAF和45DAF的种子中都有表达，且以15DAF表达量最低，随后呈上升趋势。其表达量的变化表明GsADF1通过调节肌动蛋白可能参与了种子从胚胎发生到贮藏物质积累的转变。
Abstract:
As an important kind of actin binding proteins，actin depolymerizing factor(ADF) with a low molecular weight exsist in eukaryotic cells extensively.ADF play a key roll in cell division，cell movement，plant top growth，such as pollen tube elongation and root hair formation.In this study，we took cDNA of leaves from soybean (Glycine soja) as the template，and cloned GsADF1 by RT-PCR.The full length of GsADF1 was 693 bp，which including 417bp open reading frame(ORF).GsADF1 contained 139 amino acid residues.Comparising multiple amino acid sequences between GsADF1 and ADFs protein from other species，we constructed a phylogenetic tree.We found that they were high homologous and greatly conserved in many amino acid sites.To further study the expression level of GsADF1 in different tissues and differnt development stages of soybean，we chose Real time RT-PCR to analyze it.The results indicated GsADF1expressed in roots，stems，leaves，flowers and seeds.Especially highly expressed in roots and flowers;whereas expressed in 15DAF lowly，and presented a upward trend from 15DAF to 45DAF.The change of expressing level showed GsADF1 possibly involved in the process from embryogenesis to the accumulation of storage material in the seeds.

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备注/Memo

基金项目：国家重点基础研究发展规划资助项目（2004CB117206）；国家高技术研究发展计划资助项目（2006AA10Z1C1）。

作者简介：何慧 (1981-) ，女, 硕士研究生, 研究方向为植物基因工程。E-mail:qqbal@163.com。
通讯作者：喻德跃, 教授, 博士生导师。E-mail：dyyu @njau.edu.cn。

更新日期/Last Update: 2014-09-13