转录调控基因GmLEC1的cDNA克隆及其植物表达载体的构建

doi:10.7668/hbnxb.2009.01.015

华北农学报 ›› 2009, Vol. 24 ›› Issue (1): 64-68. doi: 10.7668/hbnxb.2009.01.015

所属专题： 生物技术；

转录调控基因GmLEC1的cDNA克隆及其植物表达载体的构建

朱红林¹, 沙爱华², 符秀梅¹, 李小靖¹, 陈银华¹, 单志慧², 邱德珍², 周新安²

1 海南大学教育部热带生物资源重点实验室海南海口 570228;
2 中国农业科学院油料作物研究所湖北武汉 430062

收稿日期:2008-09-26 出版日期:2009-01-28
作者简介:朱红枚1973-)女，湖南益阳人，硕士，主要从事人豆转基因研究
基金资助:
中国农业科学院油料作物研究所基金;国家“863”计划(2006AA100104)

Molecular Cloning of Transcription Factor LECl and Constructio of Plant Expression Vector for Sense and Antil sense

ZHU Hong-lin¹, SHA Ai-hua², FU Xiu-mei¹, LI Xiao-jing¹, CHEN Yin-hua¹, SHAN Zhi-hui², QIU De-zhen², ZHOU Xin-an²

1 Key Laboratory of Tropical Biological Resources Ministry of Education Hainan University Haikou 570228 China;
2 Institute of Oil Crops Research Chinese Academy of Agriculture Sciences Wuhan 430062 China

Received:2008-09-26 Published:2009-01-28

摘要/Abstract

摘要： 以高油大豆中豆32开花后30 d的种子为材料,根据已报道的拟南芥脂肪酸合成相关转录因子LEC1序列设计简并引物,采用同源序列法从大豆种子中分离了大小为850 bp的cDNA片段,测序结果表明,该片段与拟南芥中已克隆的脂肪酸合成基因高度同源,包含完整的读码框,采用酶切连接和gateway技术构建了该基因的超量表达和RNAi植物表达载体。为借助农杆菌介导法将LEC1基因转化到大豆再生植株中,对分离的LEC1基因进行功能验证,培育高油大豆新品种奠定了基础。

关键词: LEC1基因, 基因克隆, 载体构建, 大豆

Abstract: The developing seeds of high dl soybean Zhongdou 32 were employed in this study. Degenerate primerswere designed according to the reported sequence of transcription factor LECl related to fatty acid synthesis in Arabidcp2sis.A 850 by cDNA fragment was isolated from soybean seed by homolog based candidate gene method. And RNAi vectorpHGlec was constructed by teway techndogy. The analysis of sequence revealed that the fragment was highly homolo goes with the cloned fatty acid synthesis gene in Arabidqsis, including complete reading franes. Over2expression vectorwas obtained cleavage and ligation and RNAi vector pHGlec was constructed by teway technology. This study laid afoundation for cultivating high oil soybean varieties by delivering LECI gene into soybean viaAgrobacteriu mediated andfunctional verification of LECL gene.

Key words: LECI gene, Gene cloning, Vector construction, Soybean

中图分类号:

S565

朱红林, 沙爱华, 符秀梅, 李小靖, 陈银华, 单志慧, 邱德珍, 周新安. 转录调控基因GmLEC1的cDNA克隆及其植物表达载体的构建[J]. 华北农学报, 2009, 24(1): 64-68. doi: 10.7668/hbnxb.2009.01.015.

ZHU Hong-lin, SHA Ai-hua, FU Xiu-mei, LI Xiao-jing, CHEN Yin-hua, SHAN Zhi-hui, QIU De-zhen, ZHOU Xin-an. Molecular Cloning of Transcription Factor LECl and Constructio of Plant Expression Vector for Sense and Antil sense[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(1): 64-68. doi: 10.7668/hbnxb.2009.01.015.

http://www.hbnxb.net/CN/Y2009/V24/I1/64

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转录调控基因GmLEC1的cDNA克隆及其植物表达载体的构建

Molecular Cloning of Transcription Factor LECl and Constructio of Plant Expression Vector for Sense and Antil sense

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