杭白菊CmbHLH96的克隆及生物信息分析

Cloning and bioinformatics analysis of CmbHLH 96 in‘Hangbai’

bHLH是一种转录因子,在植物生长发育过程中起着重要作用。该试验以杭白菊试管苗叶片为材料,提取RNA反转录成cDNA,设计特异引物克隆CmbHLH 96基因,其全长1203 bp,最大开放阅读框(ORF)918 bp,编码306个氨基酸序列,分子量1121.74 KD,理论等电点5.06,不稳定系数35.95,平均亲水性0.75,脂肪系数33.65。分析CmbHLH96蛋白结构域发现,其包含1个Helix-Loop-Helix结构域;对CmbHLH96蛋白二级、三级结构预测的结果它们主要由α螺旋、无规则卷曲和延伸链构成,分别占25.82%、14.71%和59.48%。亚细胞定位预测CmbHLH96可能定位在细胞核中。通过在NCBI上进行BLAST对比并构建系统进化树,结果发现CmbHLH96与青蒿同源蛋白亲缘关系比较近。miRNA预测显示CmbHLH 96基因受miR7696调控。

The bHLH is a transcription factor that plays an important role in growth and development of plants.In this experiment,we used seedling leaves of Chrysanthemum morifolium Ramat'Hangbai'as materials to extract RNA,reversely transcribed RNA into cDNA,and designed specific primers for cloning CmbHLH 96 gene.The results showed the CmbHLH 96 full length was 1203 bp,the largest open reading frame was 918 bp,and encoded 306 amino acid sequences,which the molecular weight of CmbHLH 96 was 1121.74 KD with theoretical isoelectric point of 5.06,instability coefficient of 35.95,average hydrophilicity of 0.75,and fat coefficient of 33.65.The protein domain analysis of CmbHLH96 revealed that it contained one Helix-Loop-Helix structure function domain.The secondary and tertiary structure of CmbHLH96 was predicted to be mainly composed ofα-helix,random coils,and extended chains,accounting for 25.82%,14.71%,and 59.48%,respectively.Subcellular localization was predicted that CmbHLH96 might be localized in the nucleus.Phylogenetic tree was constructed through BLAST comparison on NCBI,indicating that CmbHLH96 is closely related to the homologous protein of Artemisia annua.Prediction of miRNA showed that CmbHLH gene maybe regulated by miR7696.