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从脐橙(Citrussinensis Osbeck)果皮褐变相关基因的cDNA抑制差减文库中,筛选了一个与NAC基因家族同源的EST序列,通过RACE技术成功克隆了CsNAC基因全长cDNA序列共1203bp,并对其进行了序列分析。结果表明,CsNAC包含一个918bp的开放阅读框(ORF),编码305个氨基酸,预测的蛋白质分子量为35.2kD,等电点为6.72;CsNAC蛋白N端具有一个高度保守的NAC结构域;进化树分析表明CsNAC属于NAC蛋白家族的ATAF亚家族,可能参与胁迫反应。荧光定量PCR表达分析结果表明,CsNAC在果实贮藏过程中,随着果皮褐变的发生,与对照相比褐变果皮中的表达水平明显增强。说明CsNAC基因与脐橙果实的果皮褐变具有密切关系。
A cDNA sequence homologous to the NAC gene family was screened from cDNA subtracted cDNA library of peel browning genes in Citrus sinensis Osbeck. The full-length cDNA sequence of CsNAC gene was successfully cloned by RACE and the total length of the cDNA was 1203 bp. Sequence analysis was performed. The results showed that CsNAC contained a 918 bp open reading frame (ORF) encoding a polypeptide of 305 amino acids with a predicted molecular weight of 35.2 kD and an isoelectric point of 6.72. CsNAC protein had a highly conserved NAC domain at its N-terminus. It indicated that CsNAC belongs to ATAF subfamily of NAC protein family, which may be involved in the stress response. Fluorescent quantitative PCR analysis showed that the expression of CsNAC in the browning peel was significantly increased with the peel browning occurred during fruit storage. This indicated that the CsNAC gene is closely related to the browning of the peel of navel orange fruit.