| 付国勇，张永永，刘新春，李东梅，蔡 羽，时丽洁，王丹丹，冯宗云.青稞幼苗期叶片中 SBEⅡa基因的克隆及其表达分析[J].麦类作物学报,2019,(6):645 |
| 青稞幼苗期叶片中 SBEⅡa基因的克隆及其表达分析 |
| Cloning and Expression Analysis of SBEⅡa Gene in Hulless Barley at Seedling Stage |
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| DOI：10.7606/j.issn.1009-1041.2019.06.03 |
| 中文关键词: 青稞 淀粉分支酶 实时荧光定量PCR 酶活性 |
| 英文关键词:Qingke(hulless barley) Starch branching enzyme Real-time PCR Enzymatic activity |
| 基金项目:国家现代农业产业技术体系（大麦青稞）建设专项（CARS-05） |
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| 中文摘要: |
| 为进一步研究淀粉分支酶（Starch branching enzyme,SBE）在青稞叶片淀粉代谢中的作用，以青稞品种喜马拉雅2号和康青1号为材料，通过同源克隆技术分离 SBEⅡa基因，并对分离得到的 SBEⅡa基因进行生物信息学分析，同时利用实时荧光定量PCR（qRT-PCR）技术和淀粉分支酶活性测定试剂盒分析了在高温、低温和盐胁迫条件下 SBEⅡa基因的表达情况和SBEⅡa蛋白酶活性的变化情况。结果表明，从喜马拉雅2号和康青1号克隆得到的 SBEⅡa基因ORF区为2 466 bp，编码821个氨基酸。生物信息学分析表明，该基因编码的蛋白质分子量为92.09 kDa，预测等电点(pI)为5.4，是酸性蛋白；该蛋白酶的不稳定系数为 38.03，属于稳定蛋白；参试材料与大麦的亲缘关系最近，与细江蓠的亲缘关系最远。qRT-PCR和蛋白酶活性测定结果表明， SBEⅡa基因在不同环境下表达水平和SBEⅡa蛋白酶活性变异具有高度的正相关性，在不同环境和不同材料之间没有明显差异。 |
| 英文摘要: |
| In order to further study the role of starch branching enzyme(SBE) gene in starch synthesis pathway,hulless barley Himalaya 2 and Kangqing 1 were used to clone SBEⅡa gene bythe method of homologous cloning in this study followed by bioinformatics analysis.We got a length of open reading frame (ORF) of 2 466 bp encoding 821 amino acids.The sequence analysis showed that the emzyme SBEⅡa was a stable,hydrophobic and acid protein with the molecular weight of 92.09 kDa and theoretical pI of 5.4.It belonged to the stable protein with instable coefficient of 38.03(<40).The test materials have a high sequence homology with barely and the farthest genetic relationship with gracilaria tenuispitata.The result of real-time quantitative PCR and enzymatic activity determination showed the high positive correlation between the expression levels of SBEⅡa gene and the change trend of activity of SBEⅡa enzyme under difference kinds of stress tolerance,while there is no notable difference in different environments and materials. |
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