利用RT-PCR和RACE技术,克隆得到茶叶中合成黄酮类化合物的关键酶基因-黄酮合成酶Ⅱ,该基因属于细胞色素P450家族,在GenBank登录号为FJ169499.黄酮合成酶ⅡcDNA全长序列1 824 bp,其中1 605 bp为编码区,编码534个氨基酸,分子量为60.4 kD.通过SYBR Green I实时荧光定量PCR检测,发现黄酮合成酶Ⅱ在茶树一芽二叶春梢、当季成熟叶、花瓣、花蕊、种子中都有表达,成熟叶的表达量显著高于其他四种组织.
Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant
Using RT-PCR and RACE techniques, flavone synthase Ⅱ (FSⅡ) gene, the key enzyme in synthesize flavone, was cloned in the tea plant. The FS Ⅱ belongs to cytochrome P450 superfamily. The GenBank Accession No. is FJ169499 in the NCBI. The FS Ⅱ gene cDNA had 1 824 bp in full-length with 1 605 bp in coding region, encoding 534 amino acids. The putative molecular weight was 60.4 kD. The expression levels of flavone synthase Ⅱ gene in spring 'two and a bud', mature leaves, petals, stamen and pistils, and seeds of the tea plant were assessed using SYBR Green Ⅰ Real-time PCR. The expression in mature leaves was significantly higher than that in the other four tissues.
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