茄子生长素响应因子SmARF8 的克隆与分析.pdfVIP

中国农业科学 2009,42(7):2434-2441 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2009.07.023 茄子生长素响应因子 SmARF8 的克隆与分析 杜黎明，毛伟海，包崇来，胡天华，朱琴妹，胡海娇 （浙江省农业科学院蔬菜研究所，杭州 310021 ） 摘要： 【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3 671 bp，编码阅读框全长2 676 bp。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。 关键词：茄子；生长素响应因子；基因克隆 Cloning and Characterization of ARF Family Gene SmARF8 in Solanum melongena L. DU Li-ming, MAO Wei-hai, BAO Chong-lai, HU Tian-hua, ZHU Qin-mei, HU Hai-jiao (Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021) Abstract: 【Objective 】 Cloning of auxin response factor, SmARF8, and its expression analysis can provide a basis for molecular mechanism research of fruit development in eggplant. 【Method 】 RT-PCR and RACE were used to amplify full-length cDNA sequence of SmARF8 in eggplant. The analysis of conserved domains and phylogenetic relations among the known ARFs was performed by amino acid sequence alignment method. Real-time PCR was used to analyze the expression patterns of SmARF8 in different tissues. 【Result 】 Full-length cDNA sequence of this gene is 3 671 bp with an ORF of 2 676 bp in length. The results of phylogenetic analysis among ARF family genes indicated that this gene was highly homologous with the parthenocarpy-related gene ARF8 in Arabidopsis, so it was named SmARF8. The amino acid sequen