箭筈豌豆AGAMOUS同源基因及其启动子的克隆和分析

Isolation and Analysis of AGAMOUS Homologous Gene and Its Promoter from Vicia sativa L.

AGAMOUS(AG)是参与植物雌蕊和雄蕊发育调控的最重要花器官同源基因之一。通过TAIL-PCR、RACE和RT-PCR技术相结合,获得了箭筈豌豆花器官同源基因VsAG(VsAGAMOUS)及其上游调控序列。该基因DNA序列总长3158bp,CDS区域735bp,编码含有244个氨基酸残基的蛋白产物。序列在氨基酸水平上与近缘植物豌豆PsAG基因序列一致性达98%,与拟南芥AtAG基因一致性为68%。将该基因在GenBank上注册,登录号为JF313850。生物信息学分析表明,VsAG基因第2内含子区域含有丰富的转录调控元件,在种类和功能分化上与基因上游调控序列表现出相似特征。这一结果暗示了第2内含子对豆科植物AG基因表达调控的重要作用,并为通过基因工程方法创造箭筈豌豆优良育种材料奠定了基础。

Common vetch （Vicia sativa L.） is considered to be an idealliquidity spring proteid forage for Tibet stockbreeding. However, little has been known about the molecular mechanism of floral organ formation and subsequent seed development process during domestication of common vetch in alpine steppe. AGAMOUS（AG） gene is one of the most important floral homologous genes involved in plant stamen and gynoecium development. In this study, an AG homologue gene and its upstream regulation sequence were isolated from Vicia sativa L. using hiTAIL-PCR, RACE, and RT-PCR. The results showed that this gene was 3 158 bp in DNA sequence, coding a protein product of 244 animo acides. Homological analysis showed that the similarity between this sequence and Pisum sativum PsAG gene was over 98% in animo acid level. Therefore, the sequence was considered to be Vicia sativa AG gene. It was named VsAG with the accession number JF313850 in GenBank. By analysing the cis-elements existing in upstream promoter region and the second intron, the results showed VsAG second intron region shared some similar cis-elements with promoter region. It indicated the second intron of VsAG gene may play an important role in gene expression regulation.