巴拉圭瓜多竹CCR基因的克隆与分析

Cloning and Sequence Analysis of Cinnamoyl-CoA Reductase Gene from Guadua paraguayana

肉桂酰辅酶A还原酶(cinnamoyl-Co A reductase,CCR)是木质素生物合成途径中关键酶。本研究依据转录组数据设计特异性引物,采用RT-PCR方法成功地从巴拉圭瓜多竹(Guadua paraguayanan)中克隆得到一个全新的CCR基因的全长cDNA序列,命名为GpCCR。序列分析结果表明,GpCCR包含完整的c DNA开放阅读框(ORF),由1065bp组成,编码354个氨基酸。Blast比对结果显示该蛋白质属于CCR家族蛋白;系统进化树结果显示瓜多竹与禾本科植物大麦、水稻等亲缘关系较近。荧光定量PCR检测显示GpCCR在巴拉圭瓜多竹的茎秆中表达量最高,在叶中表达量最低。

Cinnamoyl-coA reductase （CCR） is an important enzyme in the biosynthetic pathway of lignin. GpCCR was successfully obtained from Guadua paraguayanan by RT-PCR with the primers specially designed based on the data from transcriptome. Sequence analysis showed the GpCCR had complete cDNA open readimg frame （ORF）, which consisted of 1065bp encoding 354 amino acids. Blast analysis showed that GPCCR belonged to the CCR family. Phylogenetic analysis showed that the Guadua paraguayanan had closer relative relationship with Hordeum vulgare and Oryza sativa in the Gramineae. Real time PCR detection showed GpCCR was expressed the strongest in culms and the lowest in leaves of Guadua paraguayana