摘要
为构建一新型低毒且受四环素负调控的基因表达调控系统,克隆了大肠杆菌XL1-BLUE四环素阻遏蛋白基因TetR和拟南芥热激因子基因Athsf1a缺失片段AtHSFC157,将两基因融合获得TetR:AtHSFC157融合片段作为四环素调控系统转录激活子,将其克隆入表达载体pXNSC2020中,成功构建了四环素负调控系统p35SC157-Om35Sgus。采用冻融法将其导入根癌农杆菌EHA105中,并通过农杆菌介导烟草叶片瞬间表达,以组织化学法检测GUS基因表达活性;结果在不加四环素处理条件下组织化学法检测到很强的GUS活性,而在含有1mg/L四环素的培养基中培养却没有检测到GUS活性;结论表明该技术所构建的新型四环素调控系统具有典型的受四环素负调控功能。
In order to construct a new negative regulation system which is regulated by tetracycline and lower toxin, the cloned Tetracycline Repressor Protein TetR of Ecoil. XL1-BLUE with Heat Shock Transcriptional Factor A thsfla of Arabidopsis thaliana lacking fragment AtHSFC157 by the gene fusing was recombined. Then, the expression vector-pXNSC2020 to construct the Tetracycline negative regulable system of p35SC157-Om35S gus was cloned. In the end, the plant expression vector was transformed into Agrobacterium tumefaciens EHA105 by freezing and melting, thereafter, it was taken to infect the tobacco leaf, whereafter, transient expression of the GUS gene was measured by histochemically method. The results indicated that the expression live of GUS was very strong in the condition of dealing with no Tetracycline, on the contrary, not signal in the culture medium including 1mg/L Tetracycline. It showed that the new tetracycline regulation system took on the typically negative regulation function by tetracycline.
出处
《中国农学通报》
CSCD
2007年第3期89-93,共5页
Chinese Agricultural Science Bulletin
关键词
四环素
基因表达调控系统
负调控系统
瞬间表达
Tetracycline, Gene expression regulation system, Negative regulation system, Transient expression
