CAJ | 학술논문

本研究以酿酒葡萄（Vitis vinifera）品种赤霞珠（Cabernet Sauvignon）及霞多丽（Chardonnay）为试材,采用in silico克隆和分子克隆相结合的策略,从果实中克隆到分支酸合成酶基因,命名为VvCS。该基因的cDNA编码区全长1312bp,编码436个氨基酸残基,预测其编码蛋白质分子量为46.9kD,等电点为7.8;生物信息学分析显示VvCS的DNA全长7117bp,包含13个外显子和12个内含子,定位于葡萄的第13号染色体上。VvCS编码的蛋白与其它植物来源的分支酸合成酶在氨基酸水平上的同源性为75%左右;实时荧光定量PCR分析表明VvCS在葡萄果实、茎、叶和叶柄组织中均有表达,且在果皮、果肉和种子中的表达变化趋势相似,与盛花后5周的果实相比,盛花后11周果实各部位中VvCS表达丰度均有不同程度增加。
본연구이양주포도（Vitis vinifera）품충적하주（Cabernet Sauvignon）급하다려（Chardonnay）위시재,채용in silico극륭화분자극륭상결합적책략,종과실중극륭도분지산합성매기인,명명위VvCS。해기인적cDNA편마구전장1312bp,편마436개안기산잔기,예측기편마단백질분자량위46.9kD,등전점위7.8;생물신식학분석현시VvCS적DNA전장7117bp,포함13개외현자화12개내함자,정위우포도적제13호염색체상。VvCS편마적단백여기타식물래원적분지산합성매재안기산수평상적동원성위75%좌우;실시형광정량PCR분석표명VvCS재포도과실、경、협화협병조직중균유표체,차재과피、과육화충자중적표체변화추세상사,여성화후5주적과실상비,성화후11주과실각부위중VvCS표체봉도균유불동정도증가。
A gene encoding chorismate synthase,named VvCS,was cloned from winegrape berries （Vitis vinifera） through adopting both in silico cloning and molecular cloning strategies.The full-length cDNA of VvCS contained an open reading frame of 1 312 bp in length,which encoded a polypeptide of 436 amino acid residues with a molecular mass of 46.9 kD and a pI of 7.8.Bioinformatics analysis showed that the genomic VvCS gene from genomic DNA locating on the 13th chromosome consisted of 7 117 bp in length,including 13 exons and 12 introns.The sequence homology analysis revealed that the VvCS had the homology of approximately 75% at amino acid level comparing with other plant CS proteins deposited in GenBank.Real time PCR analysis exhibited the VvCS gene transcriptionally expressed in all the tested tissues including fruits,stems,leaves and petioles,and similar trends in the transcript abundance were observed amongst the skin,pulp and seeds.Furthermore,the transcript abundance of these three tissues in the berries of 11 weeks after fully flowering showed relatively higher than that of the berries of 5 weeks after fully flowering.