摘要
目的:克隆地被菊‘神韵’和‘繁星粉’的二氢黄酮醇4-还原酶(DFR)基因,并研究其表达特性。方法 :利用RT-PCR方法克隆‘神韵’DgDFR1和‘繁星粉’DgDFR2基因,并进行生物信息学分析;通过Northern印迹检测DgDFR1和DgDFR2基因的表达特性。结果:DgDFR1和DgDFR2的开放读框分别为1125和1074 bp,分别编码由374和357个氨基酸残基构成的多肽,与菊花(Chrysanthemum×morifolium)DFR的同源性分别为99%和95%,9～330位肽段具有FR_SDR_e结构域和DFR结构域。DgDFR1和DgDFR2基因具有高度同源性,核苷酸序列在18个位点存在差异;推导的氨基酸序列的同源性为96%。Northern印迹表明,DgDFR1和DgDFR2基因在花中特异性表达。结论:克隆了‘神韵’和‘繁星粉’的DFR基因,为通过转基因技术控制地被菊花色、培育地被菊花色新品种奠定了实验基础。
Objective： To clone the dihydroflavonol 4-reductase genes（DFR） of Chrysanthemum cultivars ＇Shenyun＇ and ＇Fanxingfen＇,and to study the expression trait of the genes.Methods： DgDFR1 gene of ＇Shenyun＇ and DgDFR2 gene of ＇Fanxingfen＇ were cloned by RT-PCR method and analyzed by bioinformatics software.The expression characteristics of the genes were detected by Northern blot.Results： The open reading frame of DgDFR1 and DgDFR2 genes contained 1125 and 1074 base pairs encoding 374 and 357 amino acids respectively.Amino acid sequence analysis showed that DgDFR1 and DgDFR2 were 99% and 95% identities to DFR of Chrysanthemum×morifolium respectively.The FR_SDR_e domain and DFR domain were in the amino acid sequence from the 9th to the 330th of DgDFR1 and DgDFR2.DgDFR1 and DgDFR2 genes had high identity.The nucleotide sequence of DgDFR1 and DgDFR2 genes had 18 differences,as well as the identity of deduced amino acid sequences was 96%.Northern blot results showed that DgDFR1 and DgDFR2 genes expressed in flower specially.Conclusion： The DFR genes were successfully cloned from ＇Shenyun＇ and ＇Fanxingfen＇,which established the experiment foundation of controlling the Chrysanthemum flower color by transgenic technology and cultivating the new Chrysanthemum species with different flower color.
出处
《生物技术通讯》
CAS
2012年第1期74-79,85,共7页
Letters in Biotechnology
基金
黑龙江省博士后科研启动基金(LBH-Q08146)
中央高校基本科研业务费专项资金(DL10CA03)
关键词
地被菊
二氢黄酮醇4-还原酶
基因克隆
表达特性
Chrysanthemum×morifolium
dihydroflavonol 4-reductase
gene cloning
expression trait
