摘要
目的在红花Carthamus tinctorius中克隆调控类黄酮合成的MYB转录因子基因,通过序列及表达分析初步鉴定参与调控类黄酮合成的MYB转录因子基因。方法对已报道调控类黄酮合成MYB转录因子进行序列比对,设计简并引物克隆核心序列,采用RACE技术克隆MYB转录因子基因全长,利用生物信息学方法分析基因序列,采用半定量PCR分析基因在红花不同组织及不同花期的表达情况。结果克隆得到3个调控类黄酮合成候选MYB转录因子基因,命名为CtFRMYB1、CtFRMYB2及CtFRMYB3。序列全长分别为1 223、1 080、1 348 bp,蛋白质相对分子质量分别为17 878.15、28 766.45、27 987.89。序列分析表明3个转录因子都具DNA结合功能,属MYB转录因子家族。进化分析表明CtFRMYB1和CtFRMYB2同AtMYB12关系较近。表达分析表明CtFRMYB1和CtFRMYB2仅在花中表达且在花期3(开花第3天)表达量较高。结论成功克隆3个调控类黄酮合成候选MYB转录因子基因CtFRMYB1、CtFRMYB2及CtFRMYB3,经生物信息学及表达分析初步鉴定到2个调控红花类黄酮合成MYB转录因子基因CtFRMYB1及CtFRMYB2,为红花类黄酮合成的调控机制研究奠定基础。
Objective To clone the MYB transcription factor gene in safflower,the sequence information and gene expression were analyzed to preliminarily identify the MYB transcription factor gene involved in regulation of flavonoid biosynthesis.Methods All the sequences of MYB transcription factor gene that was reported to involved in regulation of flavonoid biosynthesis were analyzed.The degenerate primers were designed to clone the core sequence.The full length of MYB transcription factor genes were cloned by RACE.The bioinformatics were used to analyze the sequences.The expression of MYB transcription factor genes in different tissues and different developmental stages of flower were analyzed by semi-quantitative PCR.Results Three candidate MYB transcription factors were cloned,named as CtFRMYB1,CtFRMYB2 and CtFRMYB3.The full lengths of the sequence were 1 223 bp,1 080 bp and 1 348 bp,respectively.And the molecular weight were 17 878.15,28 766.45 and 27 987.89,respectively.All three transcription factors have DNA binding domain and belong to the MYB family.Homologous analysis showed that CtFRMYB1 and CtFRMYB2 were closely related to AtMYB12.Expression analysis showed that CtFRMYB1 and CtFRMYB2 were only expressed in flowers,and the expression level was higher in flowering stage 3.Conclusion Three candidate MYB transcription factor genes involved in regulation of flavonoid biosynthesis（CtFRMYB1,CtFRMYB2 and CtFRMYB30） were successfully cloned.Two MYB transcription factor genes（CtFRMYB1 and CtFRMYB2） were identified by bioinformatics and expression analysis.These result laid a foundation for the molecular mechanism analysis of flavonoid biosynthesis.
出处
《中草药》
CAS
CSCD
北大核心
2017年第21期4523-4529,共7页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(81573544)
四川省科技支撑计划(2014SZ0156)
成都中医药大学校基金(ZRYY1610
ZRQN1647)
关键词
红花
类黄酮
MYB转录因子
基因克隆
RACE
序列分析
表达分析
Carthamus tinctorius L.
flavonoids
MYB transcription factor
gene clone
RACE
sequence analysis
expression analysis
