可诱导剔除选择标记基因的pBLCK载体构建

发布时间：2018-06-28 04:14

本文选题：可诱导剔除 + 选择标记基因NPTⅡ　；参考：《核农学报》2017年10期

【摘要】：为了剔除冗余且有潜在安全问题的选择标记基因,本研究通过PCR扩增出ubiquitin启动子序列、lox P序列、Cre-GR融合基因和NPTⅡ抗性基因的编码区序列,采用酶切连接的方法将其引入载体p BRACT806,从而构建一个新的可诱导剔除选择标记基因及重组酶基因的表达载体p BLCK,测序结果显示其构建成功。该载体含有Gateway组件,可进行外源目标基因的高效重组整合;具有NPTⅡ基因,利于转基因植株的筛选;具有可诱导剔除外源基因的Cre-GR组件,诱导处于lox P位点之间的抗性标记基因NPTⅡ和Cre-GR融合基因的剔除,保留目标基因表达元件。此外,目标基因、Cre-GR融合基因和NPTⅡ基因都由ubiquitin强启动子控制,可在农作物中组成型超量表达外源基因。本研究为农作物转基因安全性研究提供了新的载体系统,具有一定的育种价值。
[Abstract]:In order to eliminate the redundant and potentially safe selection marker genes, this study amplified the sequence of ubiquitin promoter sequence, LOX P sequence, Cre-GR fusion gene and NPT II resistant gene by PCR, and introduced it into the carrier P BRACT806 by enzyme cut connection, and constructed a new inducible knockout marker gene. P BLCK, the expression vector of recombinant enzyme gene, is successfully constructed. The vector contains Gateway components, which can carry out the efficient recombination of foreign target genes, have the NPT II gene, be beneficial to the screening of transgenic plants, and have the Cre-GR components that can induce the elimination of exogenous genes and induce the resistance marker between the LOX P loci. In addition, the target gene, the Cre-GR fusion gene and the NPT II gene are all controlled by the strong promoter of the ubiquitin, and the target gene, the Cre-GR fusion gene and the NPT II gene are controlled by the ubiquitin strong promoter. This study provides a new carrier system for the research of crop transgenic safety. Breeding value.
【作者单位】：天津农学院农学与资源环境学院;天津农学院基础科学学院;
【基金】：国家级大学生创新创业训练计划项目(201610061131) 天津市高校创新团队培养计划(TD12-5017) 天津市千人计划(高粱抗性生理及遗传机理的研究)
【分类号】：Q943.2

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