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辣椒(Capsicum annuum)中一个编码果实特异性伤口刺激蛋白的基因家族的特性分析:新型转座元件家族的鉴定

Characterization of a family of genes encoding a fruit-specific wound-stimulated protein of bell pepper (Capsicum annuum): identification of a new family of transposable elements.

作者信息

Pozueta-Romero J, Klein M, Houlné G, Schantz M L, Meyer B, Schantz R

机构信息

Institut de Biologie Moléculaire des Plantes, Université Louis Pasteur, Strasbourg, France.

出版信息

Plant Mol Biol. 1995 Sep;28(6):1011-25. doi: 10.1007/BF00032663.

Abstract

Using a fruit-specific cDNA as a probe we isolated and sequenced the two corresponding homologous genes (Sn-1 and Sn-2) of the bell pepper (Capsicum annuum) genome. Both genes have a single intron and numerous unusual long inverted repeat sequences. The introns share 87% homology and Sn-2 contains one 450 bp additional sequence with structural features of a transposable element, which is highly repetitive in the bell pepper genome. Surprisingly, analysis in data banks showed that genes encoding the potato starch phosphorylase (EC 2.4.1.1) and patatin contain a similar element, named Alien, in their 5'-upstream region. Alien elements are characterized by a conserved 28 bp terminal inverted repeat (TIR), small size, high AT content, potential to form stable DNA secondary structures and they have probably been inserted in TA target sites. Interestingly, the TIR of the Alien elements shares high homology with sequences existing in the TIR of extrachromosomal linear pSKL DNA plasmid of Saccharomyces kluyveri. Northern blot analyses detected Sn-1 transcripts principally in the red fruit whereas no Sn-2 transcripts were detected in neither of the samples monitored. Western blot analyses detected a 16.8 kDa Sn protein principally in the ripe red fruit and wounded areas of green unripe fruit. A comparison of the deduced amino acid sequence of Sn-1 with protein sequences in data banks revealed a significant homology with proteins likely involved in the plant's disease resistance response. Analyses at the subcellular level showed that Sn-1 is localized in the membrane of vacuoles.

摘要

我们使用一种果实特异性的cDNA作为探针,分离并测序了甜椒(辣椒属)基因组中两个相应的同源基因(Sn-1和Sn-2)。这两个基因都有一个单一的内含子和许多不寻常的长反向重复序列。内含子具有87%的同源性,并且Sn-2包含一个450 bp的额外序列,其具有转座元件的结构特征,在甜椒基因组中高度重复。令人惊讶的是,数据库分析表明,编码马铃薯淀粉磷酸化酶(EC 2.4.1.1)和类甜蛋白的基因在其5'上游区域含有一个类似的元件,命名为Alien。Alien元件的特征是具有保守的28 bp末端反向重复序列(TIR)、小尺寸、高AT含量、形成稳定DNA二级结构的潜力,并且它们可能已插入TA靶位点。有趣的是,Alien元件的TIR与克鲁维酵母染色体外线性pSKL DNA质粒的TIR中存在的序列具有高度同源性。Northern印迹分析主要在红色果实中检测到Sn-1转录本,而在监测的任何样品中均未检测到Sn-2转录本。Western印迹分析主要在成熟的红色果实和未成熟绿色果实的伤口区域检测到一种16.8 kDa的Sn蛋白。将Sn-1推导的氨基酸序列与数据库中的蛋白质序列进行比较,发现与可能参与植物抗病反应的蛋白质具有显著同源性。亚细胞水平分析表明,Sn-1定位于液泡膜。

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