番茄NP24 基因超表达载体的构建

doi:10.11924/j.issn.1000-6850.casb15110094

中国农学通报 ›› 2016, Vol. 32 ›› Issue (16): 138-142.doi: 10.11924/j.issn.1000-6850.casb15110094

所属专题：生物技术；园艺

番茄NP24 基因超表达载体的构建

赵敬一,张喜春

(北京农学院植物科学技术学院,北京 102206）

收稿日期:2015-11-17 修回日期:2016-05-17 接受日期:2016-01-22 出版日期:2016-06-17 发布日期:2016-06-17
通讯作者: 张喜春
基金资助:
科技部“中俄主要蔬菜基因资源多样化比较研究”(2011DFR31180-3)。

Construction of Over-expression Vector NP24 Gene in Tomato

Zhao Jingyi, Zhang Xichun

(College of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206)

Received:2015-11-17 Revised:2016-05-17 Accepted:2016-01-22 Online:2016-06-17 Published:2016-06-17

摘要/Abstract

摘要： 根据NCBI上提供的NP24 基因（GenBank 登录号为543979）的序列，设计全长引物，扩增编码区cDNA，通过克隆至中间载体pMD18-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CaCl2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。

关键词: 经营需求, 经营需求, 规模经营, 农地流转, 农业政策

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.

中图分类号:

S641.2

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.

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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番茄NP24 基因超表达载体的构建

Construction of Over-expression Vector NP24 Gene in Tomato

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