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大豆C3HC4型RING锌指蛋白基因GmRZFP1克隆与表达分析
  • ISSN号:1672-1810
  • 期刊名称:植物遗传资源学报
  • 时间:0
  • 页码:343-348
  • 语言:中文
  • 分类:S668.108[农业科学—果树学;农业科学—园艺学] Q513.03[生物学—生物化学]
  • 作者机构:[1]中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部作物遗传育种重点开放实验室,北京100081, [2]新疆农业大学农学院,新疆乌鲁木齐830052, [3]西北农林科技大学农学院,陕西杨凌712100
  • 相关基金:国家高技术研究发展计划(863计划)项目(2006AA10A111);中央级公益性科研院所基本科研业务费专项;国家自然科学基金项目(30700508)
  • 相关项目:小麦HSP90在ERF介导的信号传递网络中的作用机制及其功能
中文摘要:

GmDREB3基因能提高转基因烟草和拟南芥的抗逆性。利用SiteFinding—PCR技术,从大豆品种铁丰8号基因组中分离到大豆抗逆基因GmDREB3启动子片段,长度1648bp。该片段富含A/T碱基,还含有TATA—box、低温响应元件MYC及其他顺式元件MYB、CAAT-box等。将该启动子分区段与GUS报告基因连接构建表达载体,利用基因枪法转化小麦愈伤组织,并进行干旱、高盐、低温等处理,通过组织化学染色和GUS荧光定量测定分析各区段调控元件的活性。结果表明,在干旱和低温的诱导下,该启动子能激活下游GUS基因的表达,在-285~-1117区域存在与低温和干旱应答有关的重要调控元件,在-1464~-1648区域内存在抑制启动子活性的调控元件。由此推断,在逆境条件下通过启动子区域正、负调控元件的共同作用,使GmDREB3基因的表达维持在一个恰当的水平。

英文摘要:

In order to analyze transcriptional regulative mechanism of GmDREB3 gene, the GmDREB3 promoter (1 648 bp) was isolated from soybean genome using SiteFinding-PCR method. The sequences is abundant in A/T base and predicted to contain a lot of putative cis-element, such as low-temperature responsive element (MYC), dehydration responsive element (MYB), ABA responsive element (ABRE), and so on, as well as TATA-box in biological database. To identify the key promoter regulative region controlling gene expression, GmDREB3 promoter was truncated according to the prediction of putative cis-element and inserted into the site upstream of GUS reporter gene. Then, vectors containing different length GmDREB3 promoters were transferred into wheat callus by particle bombardment, and after different stress treatments, function of these putative cis-elements was analyzed by identifying activation of GUS using histochemistry staining and GUS fluorescence intensity analysis. The results indicated that the promoters between -285 and -1 648 bp (relative to the translational start site) activated expression of GUS report gene under drought and low temperature stresses. But only the promoter region between -285 and -1 117 bp could activate expression of maximal GUS gene in wheat callus. Another promoter region between -1 117 and -1 464 bp weakened the ability for activating expression of GUS gene under drought and low-temperature stress conditions. Thus, it was suggested that there are some positive regulating motif between -285 and -1 117 bp and negative regulating motif between -1 117 and -1 464 bp of GmDREB3 promoter. As a result, the expression of GmDREB3 gene can be kept in an appropriate level response to various stresses through the regulation of both positive and negative regulating motifs.

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期刊信息
  • 《植物遗传资源学报》
  • 北大核心期刊(2011版)
  • 主管单位:中华人民共和国农业部
  • 主办单位:中国农业科学院作物科学研究所 中国农学会
  • 主编:刘旭
  • 地址:北京中关村南大街12号
  • 邮编:100081
  • 邮箱:zwyczyxb2003@163.com/sina.com/caas.cn
  • 电话:010-82105794 82105795
  • 国际标准刊号:ISSN:1672-1810
  • 国内统一刊号:ISSN:11-4996/S
  • 邮发代号:82-643
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:10278