2个丹参鲨烯合酶基因的克隆和鉴定  

Cloning and identification of two squalene synthase genes from Salvia miltiorrhiza

作　　者：马艺沔[1] 袁丽钗[1] 张林甦[1,2] 侯学敏[1,3] 卢善发[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]黔南民族医学高等专科学校,贵州都匀558003 [3]山西师范大学,山西临汾041004

出　　处：《中草药》2014年第9期1307-1312,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金青年基金(81102727);国家自然科学基金面上基金(31070534)

摘　　要：目的克隆与鉴定丹参Salvia miltiorrhiza鲨烯合酶基因。方法基于丹参基因组的基因预测结果设计引物,通过RT-PCR方法,克隆丹参鲨烯合酶基因。利用生物信息软件对基因序列进行了结构分析,对基因编码多肽进行了序列同源性比较和保守结构域分析,利用实时定量RT-PCR方法对基因进行表达模式研究。结果通过RT-PCR方法扩增得到2个丹参鲨烯合酶基因(SmSQS1和SmSQS2),它们编码的多肽具有鲨烯合酶相关结构域和保守基序。这2个基因具有不同的外显子/内含子结构特征,具有不同的组织特异性表达模式和时间表达模式。结论丹参基因组中存在2个鲨烯合酶基因,它们具有不同的基因结构特征和表达模式,可能在丹参甾体类和三萜类化合物生物合成中起不同作用。Objective To clone and identify the squalene synthase genes in Salvia miltiorrhiza. Methods The primers were designed based on the predication result of S. miltiorrhiza genome genes and two squalene synthase genes （SmSQS1 and SmSQS2） fi＇om S. miltiorrhiza were amplified via RT-PCR method. Some bioinformatic methods and soft-wares were used for the gene structure analysis, the analyses of sequence homology and protein conserved domains of the two gene encoding polypeptides were carried. Real-time quantitative PCR （RT-qPCR） method was used for the analysis of gene expression patterns. Results The two squalene synthase genes of S. miltiorrhiza were obtained by RT-PCR method. The encoded polypeptide of the two genes has the related domains and a conserved motif for squalene synthases. The two genes had the different exon/intron characteristics and the different tissue-specific and time-specific expression pattems. Conclusion There are two squalene synthase genes existing in the genome of S. miltiorrhiza, which have the different gene structures and expression pattems, and probably play the different roles in the biosynthesis process of steroids and triterpenoids in S. miltiorrhiza.

关 键 词：丹参 三萜类 鲨烯合酶 基因克隆 基因表达 

分 类 号：R282.12[医药卫生—中药学]