滁菊类黄酮3′-羟化酶基因的克隆及分子特性.pdfVIP

※生物工程 食品科学 2012, Vol. 33, No. 17 145 - 蔡 华，许雪峰，诸立新，孙艳辉，代洪亮，崔 勇 (滁州学院生物与食品工程学院，安徽 滁州 239000) ：以安徽滁州地区特有的药食两用植物资源——滁菊(Dendranthema morifolium ‘chuju’)为试材，采用同源 基因克隆方法克隆滁菊类黄酮 3 ′- 羟化酶基因(flavonoid 3 ′-hydroxylase ，f3 ′h) ，NCBI 注册号 HQ256697 。结果表 明：采自不同地点的滁菊样品均能扩增出一条长381bp 的f3 ′h 基因片段，该片段与探针序列 GU249553 第 3 外显子 对应区域的核苷酸序列同源性达 98.69%，同源区域内有5 个单核苷酸多态性位点(single nucleotide polymorphisms， SNP)，其中3 个 SNPs 导致氨基酸编码序列的改变。滁菊与葡萄属( Vitis vinifera) 、高粱属(Sorghum bicolor) 、芸 苔属(Brassica napus) 、牵牛属(Ipomoea nil Magenta) 、苹果属(Malus ×domestica) 、番薯属(Ipomoea purpurea)等物 种f3 ′h 基因的核苷酸序列和氨基酸序列同源性分别在 60%～98% 和68%～97% 之间。不同植物f3 ′h 基因的分类与物 种间的亲缘关系存在明显的相关性，菊属植物f3 ′h 基因在系统进化中处于较高位置。 ：滁菊；类黄酮；类黄酮 3 ′- 羟化酶基因；外显子 3 ；序列分析 Cloning and Molecular Characterization off3 ′h Gene in Dendranthema morifolium CAI Hua ，XU Xue-feng ，ZHU Li-xin ，SUN Yan-hui，DAI Hong-liang ，CUI Yong (School of Biological and Food Engineering, Chuzhou University, Chuzhou 239000, China) Abstract ：Dendranthema morifolium ‘chuju ’, a traditional Chinese medicine resource unique to Chuzhou, Anhui, was used to clone flavonoid 3 ′-hydroxylase (f3 ′h) gene with a registration number of HQ256697 in NCBI by homology-based cloning strategy in this study. The results showed that a separate 381 bp DNA fragment was amplified by polymerase chain reaction (PCR) from three ‘chuju’ samples collected from different places. Sequence analysis showed that the amplified DNA fragments were closely homologous to the 3rd exon off3 ′h gene in Chrysanthemum morifolium (GU249553) at a homology rate of 98.69% for nucleotide sequences. Five SNPs in the homology area were observed, and only 3 of them resulted in amino acid sequence variations. The h