| 拟南芥HIS1-3基因克隆及表达载体构建 |
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| 引用本文： | 吕申超,汪海涛,鱼斌,孟卫卫,曹树青.拟南芥HIS1-3基因克隆及表达载体构建[J].合肥工业大学学报(自然科学版),2012(1):120-123. |
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| 作者姓名： | 吕申超 汪海涛 鱼斌 孟卫卫 曹树青 |
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| 作者单位： | 合肥工业大学生物与食品工程学院 |
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| 基金项目： | 安徽高等学校省级自然科学研究重大资助项目(KJ2010ZD04);合肥工业大学学生创新基金资助项目(cxsy10096) |
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| 摘 要： | 文章提取拟南芥植株总RNA,通过RT-PCR扩增出HIS1-3基因的cDNA片段,连接到pEASY-T1Simple载体上,转化到Trans1-T1感受态细胞内,筛选阳性单克隆并抽提质粒。利用限制性内切酶Eco91Ⅰ和NcoⅠ对质粒pEASY-T1-HIS1-3和植物表达载体pCAMBIA2301进行完全双酶切,通过电转化法,将重组表达载体pCAMBIA2301-HIS1-3导入根癌农杆菌C58中以期获得携带HIS1-3基因的根瘤农杆菌株,为研究HIS1-3基因的抗逆分子机理以及遗传改良作物抗逆性奠定了基础。
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| 关 键 词： | 拟南芥 HIS1-3基因 pCAMBIA2301植物表达载体 |
Cloning of Arabidopsis HIS1-3 gene and construction of its expression vector |
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| L Shen-chao,WANG Hai-tao,YU Bin,MENG Wei-wei,CAO Shu-qing.Cloning of Arabidopsis HIS1-3 gene and construction of its expression vector[J].Journal of Hefei University of Technology(Natural Science),2012(1):120-123. |
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| Authors: | L Shen-chao WANG Hai-tao YU Bin MENG Wei-wei CAO Shu-qing |
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| Institution: | (School of Biotechnology and Food Engineering,Hefei University of Technology,Hefei 230009,China) |
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| Abstract: | The total RNA was extracted from Arabidopsis seedlings,the cDNA fragments of HIS1-3 gene were amplified by RT-PCR and lighted with pEASY-T1 Simple vector.The pEASY-T1-HIS1-3 was transformed into Trans1-T1 phage resistant chemically competent cells,the single masccline colony was screened and the plasmid was extracted.Then the restriction enzymes of Eco91 Ⅰ and Nco Ⅰ were used to completely double-digeste the pEASY-T1-HIS1-3 plasmid and the pCAMBIA2301 vector.The recombinant pCAMBIA2301-HIS1-3 vector was carried into the C58 strain of Agrobacterium tumefaciens by electrotransformation.Finally,the strains of Agrobacterium tumefaciens carrying HIS1-3 gene were obtained.The study provides the basis for studying the molecular mechanism of stress-tolerance of HIS1-3 gene and improving the stress-tolerance of crops. |
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| Keywords: | Arabidopsis thaliana HIS1-3 gene pCAMBIA2301 plant expression vector |
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