冬凌草异戊烯基焦磷酸异构酶基因(IDI)克隆与分析

Cloning and Expression of cDNA Encoding Key Enzymes(Isopentenyl Diphosphate Isomerase)in Diterpenoids Biosynthesis Pathway From Rabdosia rubescens Henrsl. Hara

以冬凌草无菌苗为试验材料,在冬凌草转录组信息数据的基础之上,采用逆转录PCR技术克隆到冬凌草二萜类合成的关键酶基因:异戊烯基焦磷酸异构酶基因(isopentenyl diphosphate isomerase,IDI),并对该基因进行了相关的生物信息学分析。结果表明:IDI cDNA基因全长1 050bp,基因开放阅读框为906bp,编码301个氨基酸,其蛋白质序列理论分子量为27.4kDa,等电点为5.06,是一种亲水性蛋白。采用实时荧光定量PCR法分析其组织表达模式,发现IDI在叶中表达量相对较高,在愈伤组织中表达量最低。

cDNAs encoding isopentenyl diphosphate isomerase were obtained from the leaves of aseptic seedlings of Rabdosia rubescens by reverse transcription PCR based on the results of transcriptome sequencing.The results showed that the full-length cDNA of IDI was 1 050 bp and contained an open reading frame（ORF）of 906 bp,encoding 301 amino acids.The theoretical molecular weight was 27.4kDa and the isoelectric point was predicted as 5.06 and it was a kind of hydrophilic protein.Real-time quantitative PCR was used to detect the relative expression levels of of IDIin different tissues of Rabdosia rubescens.And the expression of IDI was relatively high in the leaves,the lowest expression in callus.