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当前位置: 首页 > 文章 > 番荔枝节律钟输出基因AsGI的克隆、亚细胞定位与表达分析 园艺学报 2015,42 (12) 2395-2404
Position: Home > Articles > Cloning,Subcellular Localization and Expression Analysis of Circadian Clock Output Gene AsGI from Sugar Apple Acta Horticulturae Sinica 2015,42 (12) 2395-2404

番荔枝节律钟输出基因AsGI的克隆、亚细胞定位与表达分析

作  者：
刘锴栋;袁长春;黎海利;陈燕;刘金祥
单  位：
岭南师范学院生命科学与技术学院
关键词：
番荔枝;GIGANTEA;生物节律钟;亚细胞定位;表达模式
摘  要：
GIGANTEA基因在植物开花和生理节律变化过程中起着重要作用。利用同源克隆和RACE-PCR的方法获得番荔枝GIGANTEA基因的全长c DNA序列,命名为As GI,Gen Bank登录号为KR095281。序列分析表明,克隆获得的番荔枝As GI基因长为3 465 bp,编码1 154个氨基酸。氨基酸序列比对显示与葡萄和油棕的GI的相似度分别为77%和76%。构建类似蛋白系统进化树显示,番荔枝As GI与香蕉、海枣、油棕等分子进化距离较近。预测As GI蛋白为转录因子,定位在细胞核中,为非分泌性蛋白质,不具备信号肽。同时构建了该基因融合GFP的植物表达载体,通过农杆菌介导瞬时转化烟草上表皮细胞并在荧光显微镜下观察,发现在烟草上表皮细胞的细胞核中有绿色荧光。实时定量RT-PCR结果表明,在不同的器官中,As GI的表达量存在差异,在叶片、花蕾和成熟花中的表达量相对较高。As GI呈现昼夜节律表达模式,长日照下叶片中As GI在白昼的表达高于短日照处理。在4月、6月和8月,As GI的表达量相对较高。该基因可能作为番荔枝花期调控分子育种的目标基因。
译  名：
Cloning,Subcellular Localization and Expression Analysis of Circadian Clock Output Gene AsGI from Sugar Apple
作  者：
LIU Kai-dong;YUAN Chang-chun;LI Hai-li;CHEN Yan;LIU Jin-xiang;Life Science and Technology School,Lingnan Normal University;
关键词：
sugar apple;;GIGANTEA;;circadian clock;;subcellular localization;;expression pattern
摘  要：
GIGANTEA gene plays an important role in several molecular regulations in plant,such as flowering and circadian rhythm oscillation.A full-length c DNA sequence of homologous GIGANTEA gene was cloned by employing homology gene cloning and RACE-PCR from sugar apple(Annona squamosa L.),which was named as As GI(Gen Bank accession KR095281). Sequence analysis showed that the As GI gene has a 3 465 bp open reading frame(ORF)encoding 1 154 amino acids. Sequence alignment displayed that it was a similarity of 77% and 76% with GIGANTEA of Vitis vinifera,Elaeis guineensis,respectively. The phylogenetic tree showed that sugar apple As GI and Musa acuminata GIGANTEA are the closest in molecular evolution distance,followed by Phoenix dactylifera GIGANTEA and Elaeis guineensis GIGANTEA. It was speculated that As GI protein is transcription factor,located in the nucleus and it is a non-secretory protein. It do not have a signal peptide. The plant expression vector was transformed into tobacco via Agrobacterium tumefaciens mediated transformation. GFP expression was observed in the nucleus of the epidermis cells of tobacco by fluorescence microscopy. The real-time PCR results suggested that As GI gene showed a high transcription level in leaf,flower bud and mature flower. Interestingly,the expression patterns of As GI gene are circadian rhythms expression. Under long-day conditions,the expression levels of As GI gene are higher than that under short-day conditions. And the the expression levels of As GI gene are relatively high in April,June,and August. This study provides some references for further exploration of photoperiod pathway of the flower in sugar apple molecular control mechanism. Meanwhile,As GI gene could be used as the target gene of molecular breeding of flowering phase.

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