红花查耳酮异构酶基因植物表达载体的构建及拟南芥转化  

Construction of CHI gene expression vector of safflower and its transformation in Arabidopsis thaliana

作　　者：刘秀明[1,2] 李晓璇[2] 姚娜[1] 王立勇[1] 李晓薇[1,2] 董园园[1] 王南[1] 孟璐璐[3] 李海燕[1,2] 

机构地区：[1]吉林农业大学生物反应器与药物开发教育部工程研究中心,吉林长春130118 [2]吉林农业大学生命科学学院,吉林长春130118 [3]吉林省科学技术厅,吉林长春130118

出　　处：《中草药》2016年第18期3279-3283,共5页Chinese Traditional and Herbal Drugs

基　　金：国家高技术研究发展计划("863")(2011AA100606);国家自然科学基金资助(31101172;31501366);吉林省科技厅(20150623024TC-11)

摘　　要：目的构建红花查耳酮异构酶(CHI)基因的植物表达载体并在拟南芥中进行超表达验证该基因的功能。方法将已经分离的红花CHI基因作为目的基因,在其两端引入Bam H I和Eco R I酶切位点,构建含有35S启动子的植物超表达载体p BASTA-CHI,通过Flora-dip法将其转化到拟南芥中,并对转基因拟南芥T2代植株进行PCR和总黄酮量检测。结果转基因拟南芥T2代植株的PCR检测,红花CHI基因已经初步整合到拟南芥基因组中,黄酮量检测表明,转红花CHI基因的拟南芥比野生型拟南芥中的黄酮量有所提高,最高提高到2.3倍。结论成功构建了含有红花CHI基因的植物表达载体,并在拟南芥中进行超表达,获得了转基因拟南芥T2植株。Objective Plant expression vector of chalcone isomerase in safflower was built and its function was verified by overexpression of CHI in Arabidopsis thaliana. Methods CHI isolated from safflower in our previous study was introduced by Bam H I and Eco R I restriction sites and constructed into the over-expression vector p BASTA-CHI containing 35 S promoter, transformed into A. thaliana by Flora-dip method. T2 plants of transgenic A. thaliana were detected by PCR and content of total flavones. Results Plant expression vector containing the safflower CHI gene was built and over expressed in A. thaliana successfully. T2 plants of transgenic A. thaliana were obtained. Conclusion PCR of transgenic T2 in A. thaliana is detected that CHI gene of safflower has been integrated into the A. thaliana genome, flavone content is determined in leaves, and the results show that the flavone in transgenetic A. thaliana is higher than that in wild type, the highest strain increases by nearly 2.3 times.

关 键 词：红花 查耳酮异构酶 黄酮 拟南芥 PCR 

分 类 号：R282.12[医药卫生—中药学]