利用酵母双杂系统筛选与春兰CgSEP1相互作用的蛋白  

Identification of Cymbidium goeringii CgSEP1 Interacting Proteins by Yeast Two-Hybrid Screening

作　　者：刘换换 何国仁 向林 陈跃[2] 孙崇波[2] 胡凤荣[1] Liu Huanhuan;He Guoren;Xiang Lin;Chen Yue;Sun Chongbo;Hu Fengrong(Nanjing Forestry University,Nanjing,210037;Institute of Horticulture,Zhejiang Academy of Agricultural Sciences,Hangzhou,310021;College of Horticulture and Forestry,Huazhong Agricultural University,Wuhan,430070)

机构地区：[1]南京林业大学,南京210037 [2]浙江省农业科学院园艺研究所,杭州310021 [3]华中农业大学园艺林学学院,武汉430070

出　　处：《基因组学与应用生物学》2019年第2期687-693,共7页Genomics and Applied Biology

基　　金：林业公益性行业科研专项(201304117);浙江省自然科学基金(LY16C160004);国家青年自然科学基金(31400593);浙江省公益技术研究农业项目(2014C32100)共同资助

摘　　要：兰花形态优雅,花香馥郁,花形奇特,具有高度特化的花器官,观赏价值较高。有关兰花花器官形成分子机理以及基因组测序正在不断研究中。为了探讨兰花基因间相互作用情况,本研究以春兰为材料进行基因克隆,获得了18个花发育相关基因的编码序列,经测序鉴定分别属于ABCDE类MADS-box家族基因。再以其中春兰AP/AGL9组的CgSEP1为诱饵蛋白,亚克隆至pGBKT7质粒,构建诱饵载体,将18个蛋白编码亚克隆至pGADT7质粒。通过酵母双杂系统筛选到了3个与其互作的蛋白:pGADT7-AG1、p GADT7-SEP2、pGADT7-AGL6-3。结果表明,C类AG-like和E类SEP-like、AGL-like家族基因参与SEP1基因春兰花发育过程,并形成多种蛋白混合物,共同调控花瓣和萼片的形成,进一步揭示了春兰的成花机理。Orchids have elegant morphology, fragrant flowers, peculiar patterns, highly specialized flower organs,and high ornamental value. Molecular mechanism of organogenesis and genome sequencing of orchids’ flower organ are being studied continuously. In order to investigate the interaction between genes in orchids, in this study,the coding sequence of 18 genes related to flower development was obtained by cloning from Cymbidium goeringii.The genes belonging to ABCDE class MADS-box family were identified by sequencing. Then CgSEP1 of AP/AGL9 group of Cymbidium goeringii was used as bait protein and subcloned into pGBKT7 plasmid to construct bait vector. Eighteen protein-coding subclones were cloned into pGADT7 plasmid. There were three proteins by yeast two-hybrid system: pGADT7-AG1, pGADT7-SEP2, p GADT7-AGL6-3, which interacted with it. This suggestd that Class C AG-like and Class E SEP-like and AGL-like family genes participate in the development of SEP1 gene Cymbidium goeringii and form a mixture of proteins, which jointly might regulate the formation of petals and sepals, further revealing the flowering mechanism of Cymbidium goeringii as well.

关 键 词：诱饵蛋白 酵母双杂系统 花发育 

分 类 号：S682.31[农业科学—观赏园艺]