苦瓜MADS盒基因的克隆和表达研究  

Cloning and expression of a MADS box gene from bitter melon (Momordica charantia L.).

作　　者：杨满业[1] 赵茂俊[2] 徐莺[1] 唐琳[1] 白洁[1] 陈放[1] 

机构地区：[1]四川大学生命科学学院 [2]四川农业大学基础部

出　　处：《北京林业大学学报》2004年第4期30-34,共5页Journal of Beijing Forestry University

基　　金：教育部博士点基金项目"苦瓜花发育相关基因 (MADS boxGENE)的克隆及表达"( 2 0 0 2 0 610 88)的部分内容

摘　　要：根据MADS box基因保守区结构 ,设计简并性引物 ,利用 3’RACE方法 ,从苦瓜 (MomordicacharantiaL .)中分离出花特异表达基因的cDNA片段 .同时利用 5’RACE方法获得了全长cDNA ,命名为BAG .序列分析表明 ,该cDNA全长 10 0 1bp ,含一个编码 2 2 8个氨基酸的完整开放阅读框 ,5’端和 3’端非翻译区分别为 5 0、2 6 7个碱基 ,poly(A)尾巴长 2 2个核苷酸 ,具有典型的植物MADS box基因的结构 .该基因编码的蛋白质与黄瓜 (CUM10 ,CAG1) ,棉花(GHMADS 2 ) ,以及拟南芥AGL11等MADS盒基因蛋白质的同源性分别为 :95 %、93%、84 %、72 % .Southern杂交分析显示 ,在基因组中至少有两个拷贝存在 .应用RT PCR和Northern杂交结果证实 。The flower specific gene of bitter melon, named BAG (GeneBank accession number AY 178837), is cloned by the method of 3’ RACE and 5’ RACE using degenerate primer designed according to the MADS-box protein family conserved region from flower bud. A full-length cDNA of 1 001 bp contains a completed open reading frame of 228 amino acids, 5’ and 3’ nontranslated regions and a long ploy (A) tail. The predicted amino acids sequence of BAG show 95% sequence identity to MADS-box protein CUM10(Cucumis sativus), 93% to CAG1(Cucumis sativus), 84% to MADS box protein GHMADS-2(Gossipier hirsutum), 72% to MADS-box protein AGL11(Arabidopsis thaliana), respectively. A comparison between the putative translation product of BAG and the other MADS-box related proteins demonstrates that the highest conservation domain is found within the putative MADS-box DNA-binding domain. A second domain, the K-box involved in dimerization, which is conserved among the MADS-box proteins, is also found. Genomic Southern blotting analysis suggests the existence of at least two or three copies of the gene. Northern blotting and reverse transcriptase polymerase chain reaction indicates that the BAG cDNA is carpels and stamens specific in bitter melon.

关 键 词：BAG 苦瓜 MADS盒 性别决定 花 

分 类 号：S652.9[农业科学—果树学] Q78[农业科学—园艺学]