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脱落酸(ABA)在植物生长发育中起着重要的作用,9-顺式环氧类胡萝卜素双加氧酶基因(NCED)是其合成途径中的关键基因。采用逆转录PCR(RT-PCR)技术在花生种子中克隆到其同源基因AhNCED1,预测该基因开放阅读框为1 806 bp,编码601个氨基酸,蛋白质的分子质量为66.8 ku,等电点为8.49。通过生物信息学分析表明,该基因含有2种跨膜区,但均不明显;由17个丝氨酸、8个苏氨酸、5个酪氨酸参与磷酸化过程;同源比较发现,该基因与来自拟南芥、柱花草等的NCED具有较高的同源性,进化分析表明该基因与柱花草NCED的亲缘关系最近。笔者成功构建了该基因的亚细胞定位载体AhNCED1-GFP和超表达载体p Cambia2300EC-AhNCED1,为进一步研究花生中AhNCED1基因的功能奠定了基础。
Abscisic acid (ABA) plays an important role in plant growth and development. 9-cis-epoxy carotenoid dioxygenase gene (NCED) is a key gene in its synthetic pathway. The homology gene AhNCED1 was cloned from peanut seeds by reverse transcription PCR (RT-PCR). The open reading frame of this gene was predicted to be 1 806 bp, encoding 601 amino acids with a molecular mass of 66.8 ku and an isoelectric point of 8.49. Bioinformatics analysis showed that the gene contains two kinds of transmembrane regions, but none of them is obvious. 17 serine, 8 threonine and 5 tyrosine are involved in phosphorylation. The homology comparison shows that the gene NCED from Arabidopsis thaliana, Stylosanthes, etc. showed high homology, and phylogenetic analysis showed that this gene was closest to NCED. The author successfully constructed the subcellular localization vector AhNCED1-GFP and p Cambia2300EC-AhNCED1 overexpression vector, which laid the foundation for further study on the function of AhNCED1 gene in peanut.