摘要
采用RT-PCR、RACE等方法从超旱生耐盐植物梭梭（Haloxylon ammodendron）中扩增出甜菜碱合成过程中的限速酶胆碱单加氧酶（CMO）编码基因的cDNA序列（命名为HaCMO）,其开放阅读框为1 344 bp,推测的氨基酸序列全长为447个氨基酸残基.根据已获得的开放阅读框,构建重组植物表达载体pCAMBIA2300-HaCMO.HaCMO核苷酸序列与藜科几种盐生植物如菠菜（Spinacia oleracea）、山菠菜（Atriplex hortensis）、辽宁碱蓬（Suaeda liaotungensis）、盐角草（Salicornia europaea）和甜菜（Beta vulgaris）等的一致性均在74.5%以上,推导编码蛋白的氨基酸序列一致性均在73.5%以上,表明CMO编码基因在藜科植物中是一种比较保守的基因.研究结果为进一步从分子水平探明梭梭的抗旱、耐盐机制,挖掘并利用植物抗逆基因奠定了基础.
Through RT-PCR（reverse transcription polymerase chain reaction） and RACE（rapid amplification of cDNA ends）,a 1 344 bp fragment containing entire choline monooxygenase（CMO） coding region of 447 amino acids（aa） has been obtained（as named HaCMO） from extreme xerophyte and salt-tolerant plant Haloxylon ammodendron.Plant expression vector pCAMBIA2300-HaCMO with obtained fragment was constructed.Nucleotide sequence of HaCMO was similar to the corresponding fragment of CMO cDNA of several other plants,such as Spinacia oleracea,Atriplex hortensis,Suaeda liaotungensis,Salicornia europaea,Beta vulgaris,with a similarity of more than 74.5%.Encoded protein by HaCMO and CMO protein from above mentioned plants also shared more than 73.5% identity at the amino acid level.The result showed CMO gene was conserved in Chenopodiaceae.The study provided material for further exploring drought resistant and salt tolerant mechanisms of H.ammodendron in molecular level,and laid a foundation for finding and exploitation of resistant genes from stress-tolerant plants.
出处
《西北植物学报》
CAS
CSCD
北大核心
2010年第8期1514-1519,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
宁夏回族自治区科技攻关项目(KGZ-16-07-02)
关键词
梭梭
胆碱单加氧酶
基因克隆
植物表达载体
Haloxylon ammodendron
choline monooxygenase（CMO）
gene cloning
plant expression vector
