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.