莲瓣兰转录因子MADS家族DEF基因克隆及拟南芥遗传转化  

Clone Transcription Factor of DEF-Like MADS Family Gene from Cymbidium lianpan and Its Transformation in Arabidopsis thaliana

作　　者：李琼洁[1] 朱芮[2] 王小巧[1] 朱永平[3] 赵兴富[1] 肖靖译 贾茸 秦晓杰[4] 和凤美[1] 

机构地区：[1]云南农业大学园林园艺学院,昆明650201 [2]福建农林大学艺术园林学院,福州350000 [3]云南农业大学农学与生物技术学院,昆明650201 [4]云南师范大学文理学院城市学院,昆明650201

出　　处：《分子植物育种》2014年第4期760-764,共5页Molecular Plant Breeding

基　　金：云南省教育厅研究生项目(2012J076)资助

摘　　要：本研究以莲瓣兰‘玉兔彩蝶’花瓣为材料,根据GenBank中已经登录的春兰DEF基因序列(HM106982.1)设计引物,克隆DEF基因开放阅读框,构建表达载体,转化‘拟南芥’,并验证基因功能。研究结果表明:所扩增的DEF基因开放阅读框大小为669bp,编码222个氨基酸,预测蛋白质分子质量为25.560kD。该基因具有典型的植物MADS-box基因结构域,与春兰开放阅读框同源性达99%,与其它兰花同源性都在90%以上。将DEF基因构建到植物真核表达载体pCAMBIA2300中,利用农杆菌花序法转化拟南芥,经PCR分子鉴定,获得含DEF转基因拟南芥植株。For research DEF gene function, DEF gene with open reading frame from petals of Cymbidium lianpan cv.‘Yutu Caidie＇was cloned using primers which had been designed based on Cymbidium goeringii DEF gene sequences logged in GenBank（HM106982.1）, the expression vector was constructed, Arabidopsis thaliana was transformed. The results showed that DEF gene with open reading frame was 669 bp and encoded a protein of 222 amino acids and had 25.560 kD predicted protein molecules. The gene had a typical MADS-box gene structure.Homology analysis showed that DEF shared 99% similarity with Cymbidium goeringii and above 90% similarity with other orchids. Plant over expression vector was constructed by enzyme digestion of DEF gene and pCAMBIA2300 and successive ligation, and transferred into Arabidopsis thaliana by floral dip. Transgenic plants with DEF gene were obtained successfully after molecular identification.

关 键 词：莲瓣兰 MADS DEF 克隆 转化 

分 类 号：S682.31[农业科学—观赏园艺]