| 抗冻基因CBF2表达载体构建及转化紫花苜蓿的研究 |
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| 引用本文： | 刘晓静,郝凤,张德罡,毛娟,于铁峰.抗冻基因CBF2表达载体构建及转化紫花苜蓿的研究[J].草业学报,2011,20(2):193-200. |
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| 作者姓名： | 刘晓静 郝凤 张德罡 毛娟 于铁峰 |
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| 作者单位： | 甘肃农业大学草业学院,甘肃,兰州,730070;草业生态系统教育部重点实验室中-美草地畜牧业可持续发展研究中心,甘肃 兰州 730070;甘肃农业大学农学院,甘肃,兰州,730070 |
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| 基金项目： | 甘肃省农牧厅生物技术专项(GNSW-2004-07)资助 |
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| 摘 要： | 本研究以拟南芥基因组DNA为模板,用PCR方法扩增目的基因,连接到PGEM-T Easy Vector载体上构建成克隆载体T-CBF2.用BamHI和Sacl分别对克隆载体T-CBF2和植物表达载体PBIl21进行双酶切,获得目的片段和线性质粒.在T4 DNA连接酶的作用下进行定向连接,构建成植物表达载体P-T-CBF...
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| 关 键 词： | 抗冻基因CBF2 载体构建 农杆菌 紫花苜蓿 转化 |
| 收稿时间： | 1900-01-01; |
Construction of antifreeze gene CBF2 expression vector and transformation into alfalfa callus |
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| LIU Xiao-jing,HAO Feng,ZHANG De-gang,MAO Juan,YU Tie-feng.Construction of antifreeze gene CBF2 expression vector and transformation into alfalfa callus[J].Acta Prataculturae Sinica,2011,20(2):193-200. |
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| Authors: | LIU Xiao-jing HAO Feng ZHANG De-gang MAO Juan YU Tie-feng |
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| Institution: | 1,2(1.College of Grassland Science,Gansu Agricultural University,Lanzhou 730070,China;2.Key Laboratory of Grassland Ecosystem of Ministry of Education,Sino-U.S.Centers for Grazing land Ecosystem Sustainability,Lanzhou 730070,China;3.College of Agronomy,Gansu Agricultural University,Lanzhou 730070,China) |
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| Abstract: | Arabidopsis thaliana genomic DNA was selected as a template to amplify the target gene by PCR and connect it to the PGEM-T Easy Vector for construction of the T-CBF2.The target fragment and linear plasmids were obtained from the cloning vector T-CBF2 and from the plant expression vector PBI121 with dual digestion using BamHⅠand Sac I,respectively.The plant expression vector P-T-CBF2 was built through directional connections using T4 DNA ligase.PCR identification proved that the recombinant had been transferred into Agrobacterium tumefaciens and it was then introduced into Medicago sativa cv.Hetian through Agrobacterium-mediated transformation.Transfer of the target gene to alfalfa callus was successful. |
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| Keywords: | antifreeze gene CBF2 vector construction Agrobacterium tumefaciens Medicago sativa transformation |
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