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核农学报  2015 

花生FUSCA3基因的克隆与表达分析

DOI: 10.11869/j.issn.100-8551.2015.06.1044, PP. 1044-1051

Keywords: 花生,FUSCA3基因,克隆,系统发育分析,荧光定量PCR

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Abstract:

为了挖掘花生品质性状相关功能基因,本研究以花生品种花育17号为试材,根据cDNA文库中已知的FUSCA3基因全长序列设计引物,通过RT-PCR克隆到该基因,命名为AhFUSCA3。AhFUSCA3全长为1532bp,开放阅读框ORF为1134bp,对应的基因组序列3892bp,由6个外显子和5个内含子组成,内含子剪接方式符合GT-AG剪接规则。根据编码区预测AhFUSCA3编码一条378个氨基酸组成的多肽,预测分子量为42.3kD,等电点为5.58。预测该基因编码的蛋白含有B3保守结构域,可能定位于细胞核中。蛋白序列比对和进化树分析表明,花生与大豆(Glycinemax)、菜豆(Phaseolusvulgaris)、鹰嘴豆(Cicerarietinum)和苜蓿(Medicagotruncatula)等豆科植物中的FUSCA3序列相似性最高,亲缘关系最近。荧光定量PCR结果显示,AhFUSCA3基因表达量在花生荚果、种子发育的过程中呈现先增后降的趋势,推测AhFUSCA3基因在花生胚胎形成和发育过程中发挥重要作用。本研究为阐明花生胚胎发育过程和油脂代谢机制提供了理论基础,为花生品质改良育种提供了新的基因资源。

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