飞机草类黄酮3’-羟化酶基因(CoF3’H)的克隆及其在烟草中的表达

利用RT-PCR和RACE技术从飞机草中克隆得到1628 bp的类黄酮3’-羟化酶c DNA的全长序列,命名为Co F3’H,Gen Bank登录号为HQ268505.1。Co F3’H基因的编码区长度为1524 bp,编码507个氨基酸。氨基酸序列含P450蛋白结构域和半胱氨酸亚铁血红素结合域保守区(F××G×R×C×G),利用DNAMAN软件比对显示,Co F3’H与其他植物的F3’H蛋白的同源性很高。通过农杆菌介导成功地将Co F3’H基因导入烟草,T2代的转Co F3’H基因烟草植株的黄酮含量显著高于野生型。说明Co F3’H在黄酮的生物合成中起重要作用,为后续研究飞机草的化感作用及综合利用提供基础。

Chromolaena odorata Flavonoid 3’-hydroxylase Gene Cloning and Its Expression in Tobacco

A flavonoid 3’-hydroxylase gene cDNA sequence was cloned by RACE and RT-PCR techniques from Chromolaena odorata. The obtained full-length cDNA was named as CoF3’H with GenBank accession number HQ268505.1. It is 1628 bp in length, containing a 1524 bp open reading frame, encoding 507 amino acid residues. The amino acid sequence of CoF3’H contains cytochrome P450 domain and cysteineheme binding region (F××G×R×C×G). Homology analysis by DNAMAN software showed that the deduced CoF3’H protein was highly homologous to F3’H proteins from different species. CoF3’H gene was transferred into tobacco by agrobacterium-mediated genetic transformation. There was higher content of flavonvoids in T₂ transgenic tobacco than in the wild type. This result shows that CoF3’H plays an important role in flavonoid biosynthesis, and provides a basis for researches on allelopathy and comprehensive utilization of C. odorata in the future.