萼脊兰AGAMOUS(AG)基因的克隆与表达分析  

Analysis of cloning and expression analysis of AGAMOUS gene from Sedirea japonica

作　　者：蒋素华[1] 邓祖丽颖 郝平安 王默霏[1] 袁秀云[1] 崔波[1] 

机构地区：[1]郑州师范学院生物工程研究所,河南郑州450044 [2]郑州幼儿师范高等专科学校,河南郑州450000 [3]河南农业大学生命科学学院,河南郑州450000

出　　处：《河南农业大学学报》2017年第6期801-807,共7页Journal of Henan Agricultural University

基　　金：河南省高等学校重点科研项目(14B180036);郑州市科技发展计划项目(20150448);郑州市普通科技攻关项目(141PPTGG420)

摘　　要：通过对萼脊兰AGAMOUS(AG)基因进行结构、功能分析、克隆和AG基因的表达调控等方面的探究,发现,萼脊兰AG的氨基酸序列属于植物特有的MIKC型MADS-box的C类基因,该基因c DNA全长1 002 bp,包含一个702 bp的开放阅读框,该基因命名为AG(登录号KY744276),共编码233个氨基酸。蛋白序列比对和进化树分析表明,AG蛋白与蝴蝶兰蛋白一致性最高,进化距离最近。二级结构分析表明,该蛋白分子属于亲水性蛋白,其中α螺旋占48.93%,延伸链占11.59%,不规则卷曲占39.48%。实时荧光定量PCR(qRT-PCR)分析表明,AG基因在花蕾和蕊柱中表达量很高,说明AG基因的表达具有组织特异性,在ABCDE模型中属于C类基因的特征非常明显,控制着雄蕊和雌蕊的发育。The structure,function,clone of AGAMOUS(AG) gene as well as its expression regulation were studied in this paper. It's found that the amino acid sequence of AG belonged to plant type peculiar MIKC,C gene of MADS-box class. The full length c DNA of AG was 1 002 bp with an ORF of 702 bp encoding 233 putative amino acid residues(Gen Bank: KY744276). Multiple sequence alignments and phylogenetic tree analyses showed that AG was close to the phalaenopsis amabilis with an overall sequence similarity. The encoded protein was hydrophilic and contained 48. 93% of α-helical domains,11. 59% of extended strand,39. 48% of random coil. The highest expression of the gene analyzed by real-time fluorescent quantitative PCR was in bud and stigma. It indicated that AG gene expression was tissue specificity,belonged to the class C gene characteristics in ABCDE model,and controlled the development of stamen and pistil.

关 键 词：萼脊兰 AG基因克隆 RACE技术 实时荧光定量PCR 表达分析 

分 类 号：Q786[生物学—分子生物学]