文心兰一个PEBP家族基因的克隆、表达及载体构建  

Cloning,Expression and Vector Construction of A PEBP Family Gene in Oncidium

作　　者：袁秀云[1] 梁芳[1] 蒋素华[1] 王默霏[1] 刘佳[2] 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]河南农业大学生命科学学院,河南郑州450002

出　　处：《江西农业大学学报》2014年第6期1380-1386,共7页Acta Agriculturae Universitatis Jiangxiensis

基　　金：河南省科技攻关项目(092102110128);郑州市科技攻关项目(112PPTGY250-3)

摘　　要：采用RT-PCR方法从文心兰(Oncidium‘Sweet Sugar’)中克隆到一个PEBP家族基因,该基因命名为OnFT-like(登录号AHB86975)。该基因编码区531 bp,编码177个氨基酸。蛋白序列比对和进化树分析表明,On FT-like蛋白与蝴蝶兰FT蛋白的相似性为99%,进化距离最近。qRT-PCR分析表明,On FT-like在根和花器官没有检测到表达;在苗期和花蕾期的叶片和花葶中有表达,其中在花蕾期花葶中的表达量最高;在盛花期和花后期的叶片和花葶中不表达。分析认为,On FT-like基因可能在花期转变中起调控作用。将On FT-like基因连接到p CAMBIA1301载体上,成功构建了正义和反义植物表达载体,为下一步进行文心兰On FT-like基因功能鉴定及遗传改良奠定基础。A PEBP family gene was isolated from the leaf of Oncidium‘Sweet Sugar’ by using RT-PCR approaches,the gene was named OnFT-like（The accession number in GenBank：AHB86975）.The coding region of this gene was 531 bp and it encoded 177 putative amino acid residues. Multiple sequence alignments and phylogenetic tree analyses showed that OnFT-like was close to FT from phalaenopsis hybrid with 99% sequence similarity.The expression was analysed by real-time fluorescent quantitative PCR,the results showed that the expression of OnFT-like was not detected in roots and flower organs;the expression of OnFT-like was observed in both leaves and inflorescence rachis at the seedling stage and bud stage,and the highest amount of expression appeared at the bud stages;OnFT-like was not expressed in both leaves and inflorescence rachis at the full-bloom and post-flowering stages. OnFT-like was connected to pCAMBIA1301 vectors,the sense and antisense plant expression vectors were constructed successfully and can be used for research into the function of OnFT-like and genetic improvement in Oncidium.

关 键 词：文心兰 PEBP家族 实时荧光定量PCR 基因表达 载体构建 

分 类 号：Q949.718.43[生物学—植物学]