中间锦鸡儿查尔酮合成酶基因的克隆和表达分析  

Cloning and Expression Analysis of CiCHS from Caragana intermedia

作　　者：白洁[1] 杨杞[1] 郭琳[2] 王瑞刚[1] 张春林[1] 李国婧[1] 杨飞芸[3] 

机构地区：[1]内蒙古农业大学生命科学学院,呼和浩特010018 [2]内蒙古包头市种子管理站,包头014000 [3]内蒙古农业大学食品科学与工程学院,呼和浩特010018

出　　处：《基因组学与应用生物学》2016年第1期87-93,共7页Genomics and Applied Biology

基　　金：国家自然科学基金(31360169)资助

摘　　要：查尔酮合成酶是黄酮类化合物合成途径的第一个关键酶。本研究通过其它豆科植物已知的查尔酮合成酶保守序列设计简并引物,扩增得到中间锦鸡儿查尔酮合成酶基因的保守片段,经RACE技术扩增得到基因全长。对该基因gDNA全长和cDNA全长分析显示,它是由一个内含子和两个外显子构成,该基因cDNA全长为1 548 bp,其中ORF(open reading frame)为1 176 bp,5'UTR(untranslated regions)为175 bp,3'UTR为196 bp,编码391个氨基酸,推测蛋白质质量为43.03 kD,等电点为6.24,是一种两性蛋白。序列比对和系统进化分析表明,该基因属于查尔酮合成酶家族,命名为CiCHS。实时荧光定量PCR检测发现,CiCHS在紫外处理下受到诱导,0.5 h表达量最高,之后不断降低。Chalcone synthase is the first key enzyme for flavonoids synthesis.In this study, we obtained the conservative part of CHS of Caragana interrnedia through design degenerate primer by other known leguminous plants, and acquired the full-length of CiCHS by RACE （rapid amplification of cDNA）. The full-length of gDNA and cDNA of CiCHS was analyzed, the results showed that this gene had one intron and two exons. The whole cDNA length of this gene was 1 548 bp which contained ORF （1 176 bp）, 5＇UTR（175 bp） and 3＇UTR （196 bp）. The deduced protein was an amphiphilic protein which comprised 391 amino acids with a calculated molecular weight of 43.03 kD, as well as an isoelectric point of 6.24. Sequence alignment showed that this CHS protein was a member of chalcone synthase family, thus we named it CiCHS. Real-time quantitative PCR analysis showed that the transcript of CiCHS was induced by UV with the highest level at 0.5 h, deduced subsequently.

关 键 词：中间锦鸡儿 查尔酮合成酶 基因克隆 RACE 黄酮 

分 类 号：Q943.2[生物学—植物学] S567.19[农业科学—中草药栽培]