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火把梨14-3-3基因的克隆与序列分析

DOI: 10.3969/j.issn.1000-7091.2012.03.011, PP. 55-61

Keywords: 火把梨,RACE,RT-PCR,14-3-3

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Abstract:

依据火把梨(PyruspyrifoliaNakai)14-3-3蛋白的EST序列设计基因特异引物,采用快速扩增cDNA末端技术(RapidamplificationofcDNAends,RACE),从云南火把梨中克隆一个新的14-3-3基因(Pp14-3-3)的全长cDNA序列。Pp14-3-3全长cDNA为1107bp,具有84bp5’非编码区(UntranslatedRegions,UTR)、786bp开放读码框(Openreadingframe,ORF),以及237bp3’UTR,编码含261个氨基酸的蛋白质。Pp14-3-3与其他物种14-3-3蛋白在中间功能区域高度保守,包含典型的14-3-3结构域。与已知植物14-3-3s家族成员间的氨基酸序列进行聚类分析,将Pp14-3-3聚为非ε大类14-3-3。Pp14-3-3在火把梨接受光照和没有光照的果皮以及幼嫩叶片中大量表达,且表达量稳定。对Pp14-3-3的基因克隆以及表达特性进行了分析,为后期深入研究该新基因的功能奠定了基础。

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