普通烟草β-胡萝卜素羟化酶2基因克隆与生物信息学分析  

Cloning and Bioinformatic Analysis ofβ-carotene Hydroxylase 2Gene from Nicotiana tobacum

作　　者：雷波 丁福章 赵会纳 卢坤[2] 蔡凯 潘文杰 

机构地区：[1]贵州省烟草科学研究院,贵州贵阳550081 [2]西南大学农学与生物科技学院,重庆北碚400715

出　　处：《贵州农业科学》2015年第2期24-28,共5页Guizhou Agricultural Sciences

基　　金：贵州省科学技术基金项目"运用iTRAQ技术研究烤烟主要生育期烟叶中蛋白质的相对表达量"[黔科合J字顾(2013)2195];中国烟草总公司科技重大专项子课题"中间香型特色优质烟叶物质代谢基础研究"(TS-02-20110014)

摘　　要：β-胡萝卜素羟化酶(beta-carotene hydroxylase,BCH)是植物类胡萝卜素合成代谢中的关键限速酶,为获得烟草β-胡萝卜素羟化酶2(NtBCH2)基因序列,采用同源克隆法从烟草中分离NtBCH2的基因组DNA序列,基因登录号为JX101477。结果表明:1)同源克隆法从烟草中克隆出NtBCH2基因,NtBCH2基因组DNA全长2131bp,cDNA为1 083bp,含7个外显子和6个内含子。2)NtBCH2蛋白有309个氨基酸,分子量为34.79kD,具有7个糖基化位点,11个磷酸化位点,4个跨膜域。3)系统发育树与BLAST分析表明,NtBCH2是番茄的SlBCH和辣椒CaBCH2的同源基因;GENEVESTIGATOR数据库芯片结果表明,NtBCH2在幼嫩的茎和子叶中表达量最高。Beta-carotene hydroxylase is a key rate-limiting enzyme in plant metabolism of carotenoid biosynthesis.By using homologus cloning method, a BCH （NtBCH 2）gene was isolated from common tobacco K326,and the gene accession number is JX101477.Full-length of genomic DNA and cDNA is 2131 bp and 1 083 bp with 7 exons and 6 introns.The encoded NtBCH2 protein is a 309-aa polypeptide with a calculated molecular weight （Mw）of 34.79 kDa.Bioinfomatic analysis results showed that NtBCH2 possessed four transmembrane domains, seven potential glycosylation and 11 phosphorylation sites. Phylogenetic and BLAST analysis indicated that NtBCH 2 is the orthologous gene of SlBCH and CaBCH. Microarray results in GENEVESTIGATOR database revealed that NtBCH2 was expressed at the highest level in stem andcotyledon.

关 键 词：烟草 β-胡萝卜素羟化酶2基因 香气 克隆 

分 类 号：S572[农业科学—烟草工业]