| 大白菜抗软腐病基因BrWRKY33植物表达载体的构建 |
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| 引用本文： | 寇洪超,王翠花,孙玉文,徐文玲,牟晋华. 大白菜抗软腐病基因BrWRKY33植物表达载体的构建[J]. 广西农业生物科学, 2010, 0(5) |
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| 作者姓名： | 寇洪超 王翠花 孙玉文 徐文玲 牟晋华 |
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| 作者单位： | 安徽农业大学园艺学院 山东省农业科学院蔬菜研究所 |
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| 基金项目： | 国家“863”项目“抗病虫转基因大白菜育种研究”;; 山东省农业良种工程项目;; 2010年山东省农业重大应用技术创新项目共同资助 |
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| 摘 要： | 利用农杆菌侵染法获得抗软腐病转BrWRKY33基因大白菜,首先要构建植物表达载体。本实验根据BrWRKY33基因的序列及pROK2表达载体的酶切位点设计引物(FN/RN),获得BrWRKY33基因,然后将该基因连接到克隆载体pGEM-TEasy,重新构建了克隆载体T-WRKY。经测序及酶切验证,选取PCR反应中错配率最低的产物,构建了表达载体pROK2-WRKY,并将该表达载体转入到根癌农杆菌EHA105中,并对其转化子进行了鉴定。结果表明,PCR扩增获得目的基因BrWRKY33全长约1443bp,PCR及酶切结果鉴定表明克隆载体T-WRKY已经重新构建成功。表达载体的PCR及BamHⅠ和KpnⅠ双酶切鉴定表明有4个重组子表现阳性,表明目的基因已经插入到pROK2表达载体,表达载体构建成功。PCR鉴定表明表达载体pROK2-WRKY已经转入根癌农杆菌EHA105中。本实验结果将为进一步研究大白菜抗软腐病基因的转化奠定基础。
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| 关 键 词： | 大白菜 软腐病 BrWRKY33 植物表达载体 |
Construction of a Plant Expressing Vector for BrWRKY33 Gene on Resistance against Chinese Cabbage Soft Rot |
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| Kou Hongchao,Wang Cuihua,Sun Yuwen,Xu Wenling,Mu Jinhua. Construction of a Plant Expressing Vector for BrWRKY33 Gene on Resistance against Chinese Cabbage Soft Rot[J]. Journal of Guangxi Agricultural and Biological Science, 2010, 0(5) |
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| Authors: | Kou Hongchao Wang Cuihua Sun Yuwen Xu Wenling Mu Jinhua |
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| Affiliation: | 1 Department of Horticulture of Anhui Agricultural University; Hefei; 230036; 2 Institute of Vegetable Research of Shandong Academy of Agricultural Science; Jinan; 250100; |
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| Abstract: | In order to obtain transgenic Chinese cabbage of BrWRKY33 gene on resistance against soft rot by using Agrobacterium tumefacien infection process, plant expressing vector should be contructed first. In this research, we designed two primers (FN/RN) according to the gene BrWRKY33 sequence and the restriction enzyme sites of plant expression vector pROK2 to gain the gene BrWRKY33, which was connected to the clone vector pGEM-T Easy next, then reconstructed the clone vector T-WRKY. After the validation of sequ... |
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| Keywords: | Chinese cabbage （Brassica rapa. ssp. Pekinensis） Soft rot disease BrWRKY33 Plant expression vector |
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