竹叶花椒ZaHMGS基因克隆与表达分析  

Cloning and expression analysis of ZaHMGS gene from Zanthoxylum armatum

作　　者：关淑文 王毅 郝佳波 原晓龙 陆斌 李贤忠 GUAN Shuwen;WANG Yi;HAO Jiabo;YUAN Xiaolong;LU Bin;LI Xianzhong(Yunnan Academy of Forestry and Grassland Sciences,Kunming,Yunnan 650201,China;College of Forestry,Southwest Forestry University,Kunming,Yunnan 650224,China)

机构地区：[1]云南省林业和草原科学院,云南昆明650201 [2]西南林业大学林学院,云南昆明650224

出　　处：《福建农林大学学报（自然科学版）》2020年第4期467-472,共6页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：云南特色经济林新种质创新与利用(2018BB008)。

摘　　要：采用RT-PCR方法从竹叶花椒(Zanthoxylum armatum)中克隆得到一个全新的羟甲基戊二酰辅酶A合成酶(HMGS)基因的全长cDNA序列,命名为ZaHMGS.利用NCBI、ProParam、SignalP 4.1 server、DNAMAN和MEGA 7.0软件对ZaHMGS基因进行生物信息学分析,并利用荧光定量PCR检测其在嫁接树和实生树中的表达情况.结果表明:ZaHMGS包含完整的cDNA开放阅读框(OFR),由1398 bp组成,编码465个氨基酸.其蛋白的相对分子量为51359.42 u,理论等电点pI为5.93.Blast比对结果显示该蛋白质属于HMGS家族蛋白,含有由21个氨基酸组成的活性中心"GNTDIEGVDSTNACYGGTAAL"和5个参与底物催化的保守氨基酸.系统进化树结果显示竹叶花椒与克里曼丁桔(clementine mandarin,Citrus clementina)、甜橙(sweet orange,Citrus sinensis)、番木瓜(papaya,Carica papaya)等亲缘关系较近.基因表达谱表明,ZaHMGS基因在嫁接树叶、嫁接树茎、实生树叶和实生树茎中的表达量无显著性差异,在实生树各器官的表达量从高到低分别为:幼果、芽、根、茎、大果.A new full-length cDNA sequence of hydroxymethylglutaryl-CoA synthase(HMGS)gene was cloned from Zanthoxylum armatum by RT-PCR,named ZaHMGS.The bioinformatics analysis of ZaHMGS gene was performed by NCBI,ProParam,SignalP 4.1 server,DNAMAN and MEGA 7.0 software,and its expressions in grafted trees and seedling trees were detected by fluorescence quantitative PCR.The results showed that ZaHMGS contains a complete open reading frame(OFR)of 1398 bp,encoding 465 amino acids.The relative molecular weight of the protein is 51359.42 u and the theoretical isoelectric point pI is 5.93.BLAST comparison showed that the protein belongs to HMGS family proteins.It contains 21 amino acids as active center"GNTDIEGVDSTNACYGTAAL"and 5 conserved amino acids which involve in substrate catalysis.Phylogenetic tree results showed that Z.armatum has close relationships with Citrus clementina,Citrus sinensis and Carica papaya.The gene expression profiles demonstrated that there is no significant difference in ZaHMGS expression in grafted leaves and stems,and seedling leaves and stems.The expression of ZaHMGS gene peaked in young fruit,and followed by bud,root,stem and ripe fruit in a descending order.

关 键 词：竹叶花椒 羟甲基戊二酰辅酶A合成酶 基因功能分析 生物信息学分析 

分 类 号：Q943.2[生物学—植物学]