甜橙柠檬酸合酶基因的克隆及其表达分析  

Cloning and Expression Analysis of Citrate Synthase Gene in Orange

作　　者：王滕旭 李正国[1] 杨迎伍[1] 邓伟 

机构地区：[1]重庆大学生物工程学院基因工程研究中心重庆市高校功能基因与调控新技术重点实验室,重庆400030

出　　处：《中国农学通报》2010年第10期65-69,共5页Chinese Agricultural Science Bulletin

基　　金：国家支撑计划项目(2007BAD47B02-1);重庆市重大专项资助(CSTC;2007AA1018-2)

摘　　要：柠檬酸合酶与柠檬酸的积累密切相关。为了研究该基因的功能及其调控,通过设计特异性引物对甜橙柠檬酸合酶基因进行了克隆,采用半定量PCR法对哈姆林叶片、果肉和果皮中该基因的表达模式进行检测。克隆得到甜橙柠檬酸合酶基因的cDNA序列,全长1535bp,ORF区含471个氨基酸残基,序列比对显示,哈姆林甜橙柠檬酸合酶基因与其他植物的该基因高度同源。半定量PCR结果显示果肉和果皮中柠檬酸合酶基因在各个时期的表达基本没有变化,而叶片中其表达水平随果实发育成熟不断降低,与果实中柠檬酸含量变化趋势相同。说明叶片中柠檬酸合酶基因表达与果实酸含量正相关。Citrate synthase is the enzyme which is related to the accumulation of citric acid. For understanding the role and regulation of this gene, pair of specific primers was designed for the gene cloning. Semi-PCR was used to detect the expression pattern in the leaf, flesh and peel of orange. The cDNA encoding citrate synthase was cloned. It was 1 535 bp and encoded a protein with 471 amino acids. Alignment analysis showed that citrate synthase gene had high homology with other plants. The semi-PCR results showed that the expression of citrate synthase gene had no obvious change in flesh and peel during different developmental periods, while it was decreased in leaf during the development which had an identical tendency with the change of the amount of citric acid in fruit. It showed that the expression of CS gene in leaf was positively correlated with the accumulation of acid in fruit.

关 键 词：甜橙 柠檬酸合酶 基因 克隆 表达 

分 类 号：S666.4[农业科学—果树学]