| 摘要: |
| 【目的】克隆葡萄风信子MYB转录因子基因(MaMYB1)，并对其进行生物信息学和表达模式的分析。【方法】以开蓝色花的葡萄风信子品种“亚美尼亚”为材料，利用RACE技术得到其MaMYB1基因cDNA全长,对其进行生物信息学分析,利用酵母单杂交方法检测其转录激活活性，并用实时定量PCR方法分析该基因在根、茎、叶及不同发育时期花中的表达特性。【结果】通过RACE-PCR，克隆得到953 bp的MaMYB1基因，其开放阅读框长750 bp，编码249个氨基酸，推测的蛋白分子质量约为27.7 ku，理论等电点为9.3。MaMYB1基因编码的蛋白序列中具有2个典型的MYB结构域R2和R3，且在R3结构域下游含有与BHLH蛋白结合的［D/E］Lx2［R/K］x3Lx6Lx3R结构域。MaMYB1与玉米ZmP1及拟南芥MYB12的关系较近。酵母单杂交检测表明：MaMYB1具有转录激活活性。实时定量PCR分析结果表明：MaMYB1在不同组织中均有表达，以在花中表达量较高，且在花不同发育时期表达量差异显著。【结论】从葡萄风信子中克隆到1个新的MYB基因MaMYB1；推测该基因可能参与了葡萄风信子花发育过程的调控。 |
| 关键词: 葡萄风信子 MYB转录因子 基因克隆 表达分析 |
| DOI： |
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| 基金项目:国家自然科学基金项目“单子叶植物蓝色花形成关键转录因子基因克隆与功能分析”(31170652) |
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| Cloning and expression of MaMYB1 gene in Muscari botryoides |
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JIANG Ling,LIU Ya-li,LOU Qian，et al
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| Abstract: |
| 【Objective】A novel MYB transcription factor was isolated from grape hyacinths and its biological information and expression pattern were analyzed.【Method】The blue grape hyacinths variety ‘Muscari armeniacum’ was selected as material.The full-length sequence of MYB gene was obtained using RACE method,which was designated as MaMYB1,and biological information analysis was performed.The transcriptional activity of the MYB protein was detected by yeast expression system,and the expression pattern in root,stem,leaf,and flowers at different stages was analyzed by real-time PCR.【Result】The ORF of MaMYB1 was 750 bp,encoding 249 amino acids.Sequences analysis showed that the deduced protein molecular weight of the gene was 27.7 ku,the theoretical isoelectric point was 9.3,and the proteins contained two conserved MYB domains near the N-terminus and a BHLH binding motif (［D/E］Lx2［R/K］x3Lx6Lx3R) in R3 domains.Bioinformatics analysis showed that MaMYB1 had close homolog with ZmP1 from Zea mays and MYB12 from Arabidopsis thaliana.The transcriptional activation activity of MaMYB1 protein was confirmed by the yeast system.The result of real time-PCR analysis showed the gene was predominantly expressed in flower tissues and changed significantly during flower development.【Conclusion】One novel gene encoding MYB protein,MaMYB1,was isolated from grape hyacinths.Bioinformatics analysis and real-time PCR analysis showed that MaMYB1may be involved in the regulation of flower development. |
| Key words: Muscari botryoides MYB transcription factor gene cloning expression pattern |
