| 番茄NP24基因超表达载体的构建 |
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| 引用本文： | 赵敬一,张喜春. 番茄NP24基因超表达载体的构建[J]. 中国农学通报, 2016, 32(16): 138-142 |
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| 作者姓名： | 赵敬一 张喜春 |
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| 作者单位： | 北京农学院植物科学技术学院,北京农学院植物科学技术学院 |
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| 基金项目： | 科技部“中俄主要蔬菜基因资源多样化比较研究”(2011DFR31180-3)。 |
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| 摘 要： | 根据NCBI上提供的NP24基因(Gen Bank登录号为543979)的序列,设计全长引物,扩增编码区c DNA,通过克隆至中间载体p MD18-T,将类甜蛋白基因NP24插入植物表达载体PBI121,构建超表达载体PBI121-NP24,采用Ca Cl2冻融法将其转入农杆菌EHA105菌株中。酶切鉴定表明,目的基因已经正确地插入到PBI121载体中,超表达载体构建成功。菌液PCR鉴定,超表达载体已转入农杆菌中。
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| 关 键 词： | 番茄；类甜蛋白；超表达载体；农杆菌 |
| 收稿时间： | 2015-11-17 |
| 修稿时间： | 2016-05-17 |
Construction of Over-expression Vector NP24 Gene in Tomato |
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| Zhao Jingyi and Zhang Xichun. Construction of Over-expression Vector NP24 Gene in Tomato[J]. Chinese Agricultural Science Bulletin, 2016, 32(16): 138-142 |
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| Authors: | Zhao Jingyi and Zhang Xichun |
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| Affiliation: | College of Plant Science and Technology, Beijing University of Agriculture,College of Plant Science and Technology, Beijing University of Agriculture |
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| Abstract: | According to the sequence of NP24 gene (GenBank registration number 543979) in NCBI, we designed primers and amplified cDNA encoding area. Then the cDNA was cloned to intermediate carrier pMD18-T. We built the over-expression vector PBI121-NP24 by inserting the gene NP24 into plant expression vector PBI121. The over-expression vector PBI121-NP24 was transferred into agrobacterium EHA105 by the CaCl2 freeze-thaw method. Enzyme identification showed that the gene had been correctly inserted into the PBI121 carrier and the over-expression vector was built successfully. Microbial PCR identification proved that the over-expression vector had been transferred into agrobacterium. |
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| Keywords: | tomato thaumatin-like protein over-expression agrobacterium |
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