月季乙烯受体基因的全长克隆及原核表达分析  

The Full-length Cloning of Ethylene Receptor Genes and Prokaryotic Expression of RhETR3 in Roses

作　　者：唐红姝[1] 刘道凤[1] 薛璟祺[2] 欧阳琳[1] 孙翠慧[1] 马男[1] 高俊平[1] 

机构地区：[1]中国农业大学观赏园艺与园林系,北京100193 [2]中国农业科学院蔬菜花卉研究所,北京100081

出　　处：《园艺学报》2012年第12期2421-2430,共10页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31000920;30900991);农业部园艺作物生物学与种质创制重点实验室项目

摘　　要：利用已构建的月季花朵转录组数据库,筛选得到了22个与乙烯受体同源性较高的Uni Gene片段,并利用这些片段从月季花瓣中克隆得到了3个乙烯受体基因全长,分别为RhETR1、RhETR3和RhETR5。其中,RhETR1全长为2814bp,编码一个595aa的肽链;RhETR3全长为2468bp,编码一个765aa的肽链;RhETR5全长2740bp,编码一个742aa的肽链。这3个基因推定的编码蛋白分别与枣(Ziziphus jujuba)中的ZjETR1、杨树(Populus trichocarpa)中的PtETR2和砂梨(Pyrus pyrifolia)中的PpERS2同源性最高,依次为85.2%、75.8%和80.3%。蛋白同源性及结构分析表明,RhETR1和RhETR5分别与拟南芥中的ERS1和ETR1结构相似,属于第1亚家族成员,RhETR3与ERS2结构相似,属于第2亚家族成员。在花朵自然瓶插过程中,这3个基因分别表现为下调、上调和组成型表达。利用原核表达载体,选择和月季花朵开放最为相关的RhETR3,获得了和理论分子量一致的重组蛋白,表明克隆得到的RhETR3具有完整的读码框。In this paper, twenty-two UniGene fragments that shared high homology with ethylene receptor genes were obtained from previous-constructed rose flower transcriptome database. Then the full length of three genes, RhETR1, RhETR3, and RhETRS, were cloned from rose petals. The full length and encoded peptide chain ofRhETR1, RhETR3, and RhETR5 were 2 814 bp and 595 aa, 2 468 bp and 765 aa, and 2 740 bp and 742 aa, respectively. The predicted protein of the three genes shared the highest identity with ZjETR1 in Ziziphusjujuba （ 85.2% ）, PtETR2 in Populus trichocarpa （ 75.8% ）, and PpERS2 in Pyrus pyrifolia （80.3%）, respectively. Further protein homologous and construction analysis indicated that RhETR1 and RhETR5 were similar to ERS1 and ETR1 in Arabidopsis, belonging to subfamily I RhETR3 was similar to ERS2, belonging to subfamily Ⅱ. During natural vasing life, RhETR1, RhETR3,and RhETR5 showed down-regulated, up-regulated and constitutive expression, respectively. Through a prokaryotic expression system, the recombinant protein of RhETR3 was obtained, whose molecular weight was-similar to that of the theoretical ones, indicating that the cloned full-length of RhETR3 has a complete open reading frame.

关 键 词：月季 乙烯 乙烯受体 基因克隆 原核表达 

分 类 号：S685.12[农业科学—观赏园艺]