摘要
为研究萜烯合酶亚细胞定位的改变对植物萜类及其生长发育特性的影响,以青香蕉苹果(Malus domestica Borkh cv.White Winter Pearmain)果皮、菊花、酵母和拟南芥为材料,采用PCR技术分别克隆了α-法尼烯合酶基因(AFS)、Rubisco强启动子(Ru)、线粒体定位转运肽(Mit)与叶绿体定位转运肽(Pla),并利用plant CARE软件和Signal P4.1Server网站对所得启动子和转运肽序列进行了预测分析。结果表明Ru序列具备强启动子特有的TATAbox、CAATbox以及多种相关的激素响应元件,且所得线粒体和叶绿体定位转运肽也均预测到转运肽的功能。利用所得的以上序列分别构建了由Ru强启动子驱动的线粒体和叶绿体定位的AFS基因表达载体,并成功转化获得了转基因烟草植株,实时荧光定量PCR表明目的基因在转基因植物中有较高的表达,并且存在空间表达特异性。
To study the change of terpene synthase subcellular localization influenced terpenoids metabolism and plantdevelopment,we used the White winter pearmain peels,Chrysanthemum morifolium,Yeast and Arabidopsis thalianas asexperimental materials and cloned the strong promoter of alpha-Farnesene synthase,Rubisco,transit peptide ofmitochondria(mit)and transit peptide of chloroplast(pla)by RT-PCR.Then the sequence of promoter and transit peptide werepredicted by PlantCARE and SignalP4.1Server.The results showed that Ru had typical characteristics of strong promoterelements(TATAbox,CAATbox and hormone response elements)and both of mit and pla maybe have the function of transitpeptide.We have successfully constructed the alpha-Farnesene synthase gene expression vector in different subcellularlocalization driven by rubisco strong promoter,and transformed them into tobacco.Quantitative real-time PCR showed thatthe gene had a higher expression and was tissue-specific in transgenic tobacco.
作者
丁瑞瑞
王露
王茹茹
刘宇
张元湖
成妮妮
DING Rui-rui;WANG Lu;WANG Ru-ru;LIU Yu;ZHANG Yuan-hu;CHENG Ni-ni(College of Life Sciences/Shandong Agricultural University,Tai’an 271018,China;College of Life Sciences/Linyi University,Linyi 276000,China)
出处
《山东农业大学学报（自然科学版）》
CSCD
2017年第4期576-581,共6页
Journal of Shandong Agricultural University：Natural Science Edition
基金
国家自然科学基金项目(31370359)
