大豆GmWRI1a基因启动子克隆及序列分析
发布时间：2018-10-23 18:16
【摘要】：以大豆基因组文库Phytozome公布的大豆Williams82基因组序列为参考,应用Primer Premier 5.0软件设计引物,用PCR技术扩增了大豆GmWRI1a基因的启动子序列,构建了重组克隆载体pGM-TpGmWRI1a,并通过PCR扩增对阳性克隆进行鉴定送测序。克隆获得GmWRI1a基因启动子序列1 686bp,该启动子序列除含有必需的起始转录位点、TATA-box、CTTA-box外还包含多个顺式作用元件,如光应答元件、赤霉素应答元件、表达分生组织相关元件、抗旱诱导元件等。同时,构建了该启动子植物表达载体pBI-pGmWRI1a,通过PCR扩增、限制性酶切对阳性克隆进行了鉴定,为启动子的功能研究奠定基础。大豆GmWRI1a基因启动子克隆与序列分析,将为进一步研究大豆GmWRI1a基因的表达调控及其功能分析提供参考。
[Abstract]:The promoter sequence of soybean GmWRI1a gene was amplified by PCR using the primers designed by Primer Premier 5.0 software and the soybean Williams82 genome sequence published by soybean genomic library Phytozome as reference. The recombinant clone vector pGM-TpGmWRI1a, was constructed and the positive clones were identified and sequenced by PCR amplification. The promoter sequence of GmWRI1a gene, 1686 BP, was cloned and obtained. The promoter sequence contains not only the necessary initial transcription site, but also a number of cis-acting elements, such as photoresponders, gibberellin response elements, and meristem related elements. Drought-resistant induction elements, etc. At the same time, the promoter plant expression vector pBI-pGmWRI1a, was amplified by PCR and the positive clones were identified by restriction endonuclease digestion, which laid a foundation for the functional study of the promoter. Cloning and sequence analysis of soybean GmWRI1a gene promoter will provide reference for further study on the regulation and regulation of soybean GmWRI1a gene expression and functional analysis.
【作者单位】： 东北农业大学/大豆生物学教育部重点实验室/农业部东北大豆生物学与遗传育种重点实验室;
【基金】：营养功能型转基因大豆新品种培育(2016ZX08004-003) 2016年东北农业大学“学术骨干”项目
【分类号】：S565.1;Q943.2
，
本文编号：2290083
[Abstract]:The promoter sequence of soybean GmWRI1a gene was amplified by PCR using the primers designed by Primer Premier 5.0 software and the soybean Williams82 genome sequence published by soybean genomic library Phytozome as reference. The recombinant clone vector pGM-TpGmWRI1a, was constructed and the positive clones were identified and sequenced by PCR amplification. The promoter sequence of GmWRI1a gene, 1686 BP, was cloned and obtained. The promoter sequence contains not only the necessary initial transcription site, but also a number of cis-acting elements, such as photoresponders, gibberellin response elements, and meristem related elements. Drought-resistant induction elements, etc. At the same time, the promoter plant expression vector pBI-pGmWRI1a, was amplified by PCR and the positive clones were identified by restriction endonuclease digestion, which laid a foundation for the functional study of the promoter. Cloning and sequence analysis of soybean GmWRI1a gene promoter will provide reference for further study on the regulation and regulation of soybean GmWRI1a gene expression and functional analysis.
【作者单位】： 东北农业大学/大豆生物学教育部重点实验室/农业部东北大豆生物学与遗传育种重点实验室;
【基金】：营养功能型转基因大豆新品种培育(2016ZX08004-003) 2016年东北农业大学“学术骨干”项目
【分类号】：S565.1;Q943.2
，
本文编号：2290083
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