葡萄白藜芦醇合酶基因的克隆及序列分析

Cloning and Sequence Analysis of Resveratrol Synthase Gene from Vitis vinifera

目的:从葡萄中克隆白藜芦醇合酶基因vrs1并对其序列进行生物信息学分析。方法:利用葡萄总RNA为模板,采用RT-PCR技术克隆白藜芦醇合酶基因vrs1并亚克隆进T-Vector。利用生物信息学工具对其核酸和蛋白序列进行分析。结果:测序结果显示其cDNA序列全长为1 257bp,含有一个1 179bp的开放阅读框。生物信息学分析表明葡萄白藜芦醇合酶基因编码392个氨基酸,分子量为42.9kDa,理论等电点为5.97,具有芪合酶家族固有的氨基酸保守结构域,二级结构主要由α-螺旋、无规则卷曲、延伸链和β-转角组成。结论:该基因的克隆、生物信息学分析为进一步研究其功能奠定了基础。

Objective： Resveratrol synthase gene vrs1 from Vitis vinifera was cloned and its sequence had been analyzed by bioinformatics.Method：Total RNA from Vitis vinifera was used as the template to clone the cDNA sequence of vrs1 by RT-PCR and the vrs1 fragment was subcloned into T-Vector.Bioinformatics softwares were used to analysis the cDNA and protein sequence.Result： The result of sequencing showed that the full length of cDNA is 1 257bp.The gene vrs1 contains an opening reading frame of 1 179bp,which coded for 392 amino acid residues,with molecular weight of 42.9kDa,theoretical PI of 5.97.Detailed analysis of the amino acid sequence reveal a conserved domain which found in all stilbene synthase sequences.The secondary structure of resveratrol synthase contain α-helix,β-corner,extending and random curling.Conclusion： The cloning and bioinformatic analysis of vrs1 from Vitis vinifera cv.Kyoho could provide the molecular base for further study of its biological functions.