高丛蓝莓类黄酮3′-羟化酶基因F3′H的克隆及表达特性分析  

Cloning and Expression Characteristics Analysis of Flavonoid3′-hydroxylase(F3′H) in Highbush Blueberry

作　　者：李艳芳[1] 张凌云[1] 

机构地区：[1]北京林业大学森林培育与保护教育部重点实验室,中国北京100083

出　　处：《生命科学研究》2015年第4期303-309,共7页Life Science Research

基　　金：教育部全国百篇优秀博士学位论文基金资助项目(200771)

摘　　要：以高丛蓝莓(Highbush blueberry)"喜来(Sierra)"和"日出(Sunrise)"为研究材料,通过RACE(rapid amplification of c DNA end,RACE)技术获得蓝莓(Vaccinium ssp.)F3′H基因的全长c DNA序列,并使用生物信息学工具对该基因编码蛋白的氨基酸组成及理化性质、二级和三级结构,以及氨基酸序列的相似性等进行了预测和分析。利用荧光定量PCR(RT-q PCR)手段来检测其在蓝莓果实不同发育时期表达情况。结果表明:Vs F3′H全长c DNA为1 871 bp,编码530个氨基酸,蛋白相对分子质量为58.33 k D,理论p I值为7.32;蛋白疏水性指数为0.004,属于疏水性蛋白,二三级结构预测可知其富含α-螺旋和无规则卷曲。RT-q PCR发现Vs F3′H主要在果实发育前期表达量较高,蓝果期表达量明显下调,且在不同品种中以蓝果期的表达量差异显著。这些研究结果为解析蓝莓果实色泽发育及果树分子育种奠定了理论基础。The Highbush blueberry ＂Sierra＂and ＂Sunrise＂were selected as experimental material. Fulllength c DNA sequence of Vs F3′ H was acquired by RACE（rapid amplification of c DNA end） assays. The amino acid composition, physical and chemical properties, phylogenetic tree, secondary and tertiary structure of amino acid were analyzed and predicted by bioinformatics tools. The relative expression level of Vs F3′H at different fruit growth stages using quantitative real time PCR（RT-q PCR） was measured. The results showed that the full length c DNA of Vs F3′H was 1 871 bp encoding 530 amino acids. Its theoretical molecular weight was 58.33 k D and the theoretical p I was 7.32. Protein hydrophobic index was 0.004 and it is a hydrophobic protein. The prediction of secondary and tertiary structure showed that the protein was rich inα-helix and random coil. The expression level of Vs F3′H has a significant difference at blue fruit period in different varieties. Vs F3′ H is mainly involved in fruit development at early stage and declined with color development. The differential expression of Vs F3′ H at late developmental stage of fruit may lead to the dif ferent concentrations of anthocyanin in different blueberry varieties. The aim of this study is to lay a theoretical foundation for fruit color and molecular breeding of blueberry.

关 键 词：蓝莓 类黄酮3′-羟化酶 CDNA末端快速扩增技术 生物信息学 组织表达 

分 类 号：Q781[生物学—分子生物学]