博落回bHLH转录因子的克隆、表达分析及植物表达载体构建  

Cloning and Expression Analysis of b HLH Transcription Factor from Macleaya cordata and Construction of Plant Expression Vector

作　　者：黄鹏[1,2] 刘金凤[1,2] 柳亦松[2,3] 唐其[1,2] 曾建国[1,2,3] 

机构地区：[1]湖南农业大学园艺园林学院,长沙410128 [2]湖南农业大学兽用中药资源与中兽药创制国家地方联合工程研究中心,长沙410128 [3]湖南农业大学动物医学院,长沙410128

出　　处：《分子植物育种》2017年第4期1332-1337,共6页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31200615);国家重点实验室培育基地开放课题(15KFXM09)共同资助

摘　　要：碱性螺旋-环-螺旋(bHLH)类转录因子在植物的发育、抗逆和次生代谢产物的调控等方面起重要的作用。通过将其他物种的bHLH基因序列在博落回转录组数据库中进行比对筛选到1个同源性较高的unigene(McbHLH1)并对序列生物信息学分析,该基因开放阅读框(ORF)长为744 bp,编码247个氨基酸,预测其编码的分子质量为27.021 kD,理论等电点为10.08。实时荧光定量结果显示McbHLH1基因在根部表达量最高,茎和叶子中表达量极低。表明McbHLH1可能参与博落回苄基异喹啉生物碱(BIAs)的合成与调控,并构建植物表达载体用于基因功能验证。The basic helix-loop-helix（bHLH） transcription factor plays important roles in plant development,stress resistance and regulator of secondary metabolic.We download the b HLH gene from other species and compared the sequence with M.cordata transcriptome database and screened a candidate bHLH1 gene from M.cordata.And then we used bioinformatics tools to analyze the sequence.The open reading frame（ORF） was 744 bp which encoding 247 amino acids with a predicted molecular mass of 27.021 kD and theoretical isoelectric points（pI） of 10.08.The results of fluorescence quantitative PCR indicated that the gene expression was highest in roots and very low in stems and leaves.It suggested that Mcb HLH1 played a role in the synthetic and regulator of BIAs and constructed the plant expression vector for gene functional verification.

关 键 词：博落回 bHLH1转录因子 实时定量表达 表达载体构建 

分 类 号：Q943.2[生物学—植物学] S567.239[农业科学—中草药栽培]