浙贝母HMGR基因保守区序列的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of Conserved Fragment of HMGR Gene in Fritillaria thunbergii

作　　者：冯亚斌[1] 俞信光[1] 庄欣晨 王忠华[1] 

机构地区：[1]浙江万里学院生物技术研究所,浙江宁波315100

出　　处：《核农学报》2016年第12期2289-2294,共6页Journal of Nuclear Agricultural Sciences

基　　金：浙江省科技厅育种专项(2012C12912);宁波市公益类重大专项(2015C110007);浙江万里学院重中之重学科开放基金项目(KF2015001)

摘　　要：3-羟基-3-甲基戊二酰辅酶A还原酶（HMGR）是甲羟戊酸途径中的第一个限速酶,在植物生物碱类物质代谢过程中发挥重要的作用。为探究HMGR酶在浙贝母甾体类生物碱合成代谢途径中的调控作用,以百合科浙贝母狭叶品种为材料,利用RT-PCR技术和Ta克隆技术首次从浙贝母中克隆出HMGR基因保守区序列,长度为390 bp。序列分析表明,浙贝母HMGR基因保守区序列与其他9种植物的HMGR基因有较高的相似性（84%-79%）;蛋白质同源比对、特征区及系统进化树分析表明,浙贝母HMGR氨基酸片段与其他植物有较高的同源性（95%-88%）,与球药隔重楼和海枣等单子叶植物的亲缘关系较近,据此,初步确定该基因为浙贝母HMGR基因,命名为Ft HMGR。Ft HMGR基因的克隆及分子鉴定为进一步解析该基因在浙贝母甾体类生物碱合成代谢途径中的分子调控作用奠定了基础,有助于对浙贝母活性物质的合成进行调控,从而提高产量。HMGR is the first limit enzyme of mevalonate pathway,which play an important role in steroid alkaloid biosynthesis. In order to explore the regulation of HMGR in steroidal alkaloid syntheses in Fritillaria thunbergii,RT-PCR and TA cloning technology was used in this study and the first conserved fragment of HMGR gene in Fritillaria thunbergii,which was named Ft HMGR,has been cloned. The total length of this fragment was 390 bp. Sequence analysis showed that Ft HMGR had high identity（ 79% to 84%） to nine HMGR genes from various plants,the deduced amino acid sequence had a higher homology with other plants（ 95% to 88%） and was closely related to that of Paris farqesii,Phoenix dectylifera and etc monocots. Cloning and molecular characterization of Ft HMGR would be useful for further analysis of molecular regulation of steroid alkaloid biosynthesis in Fritillaria thunbergii.

关 键 词：浙贝母 HMGR 基因克隆 生物信息学 

分 类 号：S567.231[农业科学—中草药栽培]