文心兰ACC氧化酶基因OnACO1克隆与表达分析  

Cloning and Expression Analysis of OnACO1 Gene in Oncidium

作　　者：杨光华[1,2,3] 刘进平[1,2] 

机构地区：[1]海南大学热带作物种质资源保护与开发利用教育部重点实验室 [2]海南大学农学院,海南海口570228 [3]三亚市南繁科学技术研究院,海南三亚572000

出　　处：《热带作物学报》2014年第4期693-699,共7页Chinese Journal of Tropical Crops

基　　金：海南大学校基金项目(No.2012hckled-7)

摘　　要：以文心兰切花品种"黄金2号"(Oncidium Gower Ramsey‘Gold2’)为材料,通过RT-PCR和RACE技术,从文心兰的花中分离得到了1个ACO基因成员的cDNA,命名为OnACO1(GeneBank登录号为JQ822088)。该基因全长为1 424 bp,包括972 bp的ORF,172 bp的5′-UTR和280 bp的3′-UTR,编码324个氨基酸,其氨基酸序列与石斛兰等兰科植物的ACO氨基酸序列有较高的同源性。构建原核表达载体pGEX-OnACO1,转化大肠杆菌BL21进行诱导表达,在SDS-PAGE中显示出了特异性表达条带,与预测的蛋白大小一致。荧光定量PCR分析表明OnACO1基因的表达具有组织特异性,主要在蕊柱中表达,并且乙烯对该基因的表达有上调作用。A member of ACO gene family, OnACO1 （GeneBank accession number JQ822088） was cloned from Oncidium Gower Ramsey ＇Gold2＇ by RT-PCR and RACE technology. The full length cDNA is 1 424 bp with an ORF of 972 bp encoding 324 amino acids, a 172 bp 5＇-UTR and a 280 bp 3＇-UTR. The deduced amino acid sequence of OnACO1 shares high homology with those of other orchid species. Vector pGEX-OnACO1 was constructed and transformed into E. coli BL21. Prokaryotic expression and SDS-PAGE indicated that a specific band of OnACO1 expression was in consistent with the predicted protein size. Quantitative PCR analysis showed there was a tissue-specific pattern of OnACO1 expression, mainly in gynostemium, and the expression was upregulated by ethylene.

关 键 词：文心兰 OnACO1 RACE 基因克隆 表达分析 

分 类 号：Q75[生物学—分子生物学]