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【目的】克隆茄子生长素响应因子新基因,为研究茄子果实形成发育的分子机制提供依据。【方法】采用RT-PCR和RACE技术克隆茄子ARF家族相应基因的cDNA全长序列。经序列联配进行与其它物种同源基因的保守域和进化关系分析。实时定量PCR检测目的基因在茄子不同组织中的表达情况。【结果】将该基因命名为SmARF8,它的cDNA序列全长为3671bp,编码阅读框全长2676bp。氨基酸序列分析表明,SmARF8蛋白拥有核定位序列,蛋白质结构具有ARF家族的N-端DNA结合域,中间脯氨酸、丝氨酸和苏氨酸残基富集的非保守调控域,C-末端蛋白互作域。氨基酸与拟南芥的调控单性结实基因ARF8相似性较高,进化树分析表明它们聚集在一个分支上,进化关系非常密切。SmARF8基因在幼果中表达最强,在根、茎、叶、花蕾、花朵及成熟果实中表达较强烈。【结论】从茄子中克隆得到一个生长素响应因子家族基因SmARF8。
【Objective】 The aim of this study was to clone a new gene of auxin response factor of eggplant and provide basis for studying the molecular mechanism of eggplant fruit development. 【Method】 The full-length cDNA sequence of the corresponding gene of ARF family in eggplant was cloned by RT-PCR and RACE techniques. The conserved domains and evolutionary relationships of homologous genes with other species were analyzed by sequence alignment. Real-time quantitative PCR was used to detect the expression of the target gene in different tissues of eggplant. 【Result】 The gene was named SmARF8. The full-length cDNA of SmARF8 was 3671bp and the full-length coding sequence was 2676bp. Amino acid sequence analysis showed that the SmARF8 protein has a nuclear localization sequence, the protein structure has the N-terminal DNA binding domain of the ARF family, the non-conservative regulatory domain in which the middle proline, serine and threonine residues are enriched, the C-terminal protein For the domain. Amino acids are highly similar to ARF8, a regulator of Arabidopsis thaliana, and phylogenetic tree analysis suggests that they are clustered in one branch and have a very close evolutionary relationship. The SmARF8 gene is most strongly expressed in young fruits and is strongly expressed in roots, stems, leaves, buds, flowers and mature fruits. 【Conclusion】 SmARF8, an auxin response gene family gene, was cloned from eggplant.