菘蓝莽草酸羟基肉桂酸酰转移酶IiHCT基因的克隆和表达分析

Cloning and expression analysis of a hydroxycinnamoyl-Co A shikimate / quinate hydroxycinnamoyl transferasegene( Ii HCT) from Isatis indigotica

根据菘蓝转录组数据,克隆了菘蓝莽草酸羟基肉桂酸酰转移酶Ii HCT的全长c DNA并进行生物信息学分析,进一步利用实时荧光定量PCR技术分析了Ii HCT基因的表达模式。Ii HCT ORF为1 290 bp,编码430个氨基酸,等电点为5.77,相对分子质量为47.68 k Da。Ii HCT主要在菘蓝茎中表达,幼根、叶、花蕾中几乎不表达。同时发现离体菘蓝毛状根中可以检测到Ii HCT的表达,外源茉莉酸甲酯(Me JA)处理后,Ii HCT相对表达量明显增加,在4 h达到原先的4.3倍。研究首次从菘蓝中克隆得到HCT基因,为进一步解析菘蓝苯丙素类成分的生物合成途径奠定基础。

Based on the transcriptome data,we cloned the open reading frame of Ii HCT gene from Isatis indigotica,and then performed bioinformatic analysis of the sequence. Further,we detected expression pattern in specific organs and hairy roots treated methyl jasmonate（ Me JA） by RT-PCR. The Ii HCT gene contains a 1 290 bp open reading frame（ ORF） encoding a polypeptide of 430 amino acids. The predicted isoelectric point（ p I） was 5. 7,a calculated molecular weight was about 47. 68 k Da. Ii HCT was mainly expressed in stem and undetectable in young root,leaf and flower bud. After the treatment of Me JA,the relative expression level of Ii HCT increased rapidly. The expression level of Ii HCT was the highest at 4 h and maintained two fold to control during 24 h. In this study,cloning of Ii HCT laid the foundation for illustrating the biosynthesis mechanism of phenylpropanoids in I. indigotica.