番茄果实发育早期差异表达基因的鉴定。编码RAD23蛋白的新cDNA的鉴定

M. Lemaire-Chamley, Johann Petit, M. Causse, P. Raymond, C. Chevalier
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引用次数: 6

摘要

在成熟开始之前,番茄果实的发育包括三个不同的阶段:坐果期、细胞分裂期和细胞膨大期。在这项研究中,我们使用mRNA差异显示的方法来分离在果实发育早期特异性表达的番茄基因。在40个差异条带中,选择9个经过核苷酸测序鉴定的cdna进行进一步研究。我们分离出了其中一个片段对应的全长cDNA,该片段编码RAD23,这是一种参与切除修复系统的蛋白质,从而为植物中一个特征不明确的蛋白质提供了新的序列信息。利用northern blot和半定量RT-PCR技术对分离得到的cdna在番茄果实发育早期和营养器官中的表达进行了研究。序列数据保存在GenBank中,登录号为AJ270956 (mo5-3C11/1)、AJ270957 (mo5-3G12/4)、AJ270958 (mo5-3G17)、AJ270959 (mo5-3T12)、AJ270960 (mo1-6A1)、AJ270961 (mo1-6T1)、AJ270962 (mo5-10G1)、AJ270963 (mo6-20G1)、AJ270964 (mo6-MGT2)和AJ243875 (LeRAD23-8全长cDNA)。
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Identification of differentially expressed genes during early development of tomato fruit. Characterisation of a novel cDNA coding for a RAD23 protein
Before the onset of ripening, tomato fruit development comprises three distinct phases: fruit set, a cell division phase and a cell expansion phase. In this study, we used the method of mRNA differential display in order to isolate tomato genes specifically expressed during these early phases of fruit development. Among 40 differen-tial bands, nine cDNAs were selected for further investigations based on their identification after nucleotide sequencing. We isolated the full-length cDNA corresponding to one of these fragments, coding for RAD23, a protein involved in the excision repair system, thus providing new sequence information on a poorly characterised protein in plants. All the isolated cDNAs were mapped on the tomato genome and their expression studied by northern blot and semi-quantitative RT–PCR during early fruit development and in vegetative organs of tomato plants. The sequence data are deposited in the GenBank under the accession numbers: AJ270956 (mo5-3C11/1), AJ270957 (mo5-3G12/4), AJ270958 (mo5-3G17), AJ270959 (mo5-3T12), AJ270960 (mo1-6A1), AJ270961 (mo1-6T1), AJ270962 (mo5-10G1), AJ270963 (mo6-20G1), AJ270964 (mo6-MGT2) and AJ243875 (LeRAD23-8 full-length cDNA).
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