金针菇异戊烯基焦磷酸异构酶基因的克隆及表达分析

Cloning and Expression of an Isopentenyl Diphosphate Isomerase Gene from Flammulina velutipes

PCR扩增了金针菇（Flammulina velutipes）异戊烯基焦磷酸异构酶基因（FvIDI）的DNA全长和开放阅读框序列。该基因的ORF全长753bp,DNA全长923bp,含有3个内含子和4个外显子,编码250个氨基酸,金针菇异戊烯基焦磷酸异构酶相对分子质量为28450,等电点为5.05。采用实时荧光定量PCR技术研究FvIDI基因在不同发育时期和不同温度下的表达量,结果显示,FvIDI基因在采收期菌盖中表达量最高,4℃诱导处理2~4h和37℃诱导处理2~6h的表达量明显高于常规温度21℃处理。

The full-length DNA and open reading frame （ORF） sequences of an isopentenyl diphosphate isomerase （IDI） gene were cloned from Flammulina velutipes by PCR amplification. ID1 was 923 bp in length, with an ORF of 753 bp encoding 250 amino acids, and contained three introns and four exons. IDI protein had a molecular mass of 28450 and a pI value of 5.57. Real-time quantitative PCR revealed that IDI expression levels were higher in mature pilei compared with the mycelial, primordium, stipe elongation, pileus elongation and stipe maturation stages. Expression levels were also higher in F. velutipes mycelia grown at 21℃ for 6 d before exposure to 4℃ for 2-4 h or to 37℃for 2-6 h compared with controls maintained at 21℃.