拟南芥CBF1基因植物表达载体构建及其野生蕉的遗传转化
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国家自然科学基金(31000903);广东省自然科学基金(10451064001006326);广东省农业科学院院长基金(20090104)


Construction of Plant Expression Vector of CBF1 and its Genetic Transformation of Wild Banana (Musa intinerans Cheesm)
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    摘要:

    从拟南芥中克隆CBF1基因,转入到植物表达载体pBI121的XbaI和SacI位点,得到中间重组质粒pBI121-CBF1,用EcoRI和HindⅢ将35S启动子与CBF1基因从pBI121-CBF1切下,转入到植物表达载体pCAMBIA1301中,构建植物表达载体p1301-CBF1。将构建的植物表达载体转入农杆菌EHA105,采用农杆菌介导法转化野生蕉胚性悬浮细胞,经GUS组织化学染色和PCR鉴定,证明CBF1基因已转入野生蕉中。。

    Abstract:

    The CBF1 gene(CRT/DRE binding factor 1) was obtained from Arabidopsis thaliana by PCR and cloned into pMD18-T. Then CBF1 gene was sub-cloned into pBI121 vector to abtain the middle recombinant plasmid pBI121-CBF1 via XbaⅠand Sac I restriction enzyme sites. After that, the fragments containing 35S-CBF1 were digested from recombinant plasmid pBI121-CBF1 by EcoR I and Hind III, and directionally linked to the plant expression vector pCAMBIA1301 to get the plant expression vector p1301-CBF1. We transformed the embryogenic cell suspensions of wild banana(Musa intinerans Cheesm) with p1301-CBF1 via Agrobacterium-mediated transformation. Results from histochemical GUS assay and PCR suggested that CBF1 gene has been transferred into the wild banana.

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刘凯,胡春华,魏岳荣,易干军,邵秀红.拟南芥CBF1基因植物表达载体构建及其野生蕉的遗传转化[J].湖南农业大学学报(自然科学版),2011,37(3):248-252.

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  • 收稿日期:2010-11-22
  • 最后修改日期:2011-04-06
  • 录用日期:2011-05-20
  • 在线发布日期: 2011-06-05
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