葡萄白藜芦醇合酶基因的克隆与生物信息学分析

以葡萄总RNA为模板,利用RT-PCR的方法从葡萄中扩增获得一条白藜芦醇合酶基因完整的c DNA序列,命名为RS。利用生物信息学软件对其核酸和蛋白质序列进行分析,结果表明,该序列长1179 bp,与已报道的葡萄白藜芦醇合酶基因的序列相似性达到94%⁹9%,氨基酸序列相似性为96%⁹9%;RS基因编码392个氨基酸,氨基酸序列含有完整的芪合酶家族的特征序列GVLFGPGLT和活性中心序列GCYAGGTVLR;预测的分子量为42.78 k Da,理论等电点为6.57,不稳定参数为35.92,在分类上属于稳定性蛋白;二级结构主要以α-螺旋、无规则卷曲以及β-折叠为主,其中α-螺旋含量为44.13%、无规则卷曲含量为26.53%,β-折叠含量为17.66%。 

Cloning and sequence analysis of a resveratrol synthase gene from Vitis vinifera

A complete c DNA sequence of resveratrol synthase gene was obtained from Vitis vinifera total RNA by RT-PCR method, named RS.The results of multiple sequence alignment showed that RS was of 1179 bp with 94% ~ 99% nucleic acid sequence similarity and 96% ~ 99% amino acid sequence similarity to the reported Vitis vinifera resveratrol synthase, respectively.Domain analysis results revealed that the characteristics sequences GVLFGPGLT and the active site sequences GCYAGGTVLR of stilbene synthase family were found in cloned RS amino acids sequence.The predicted molecular weight was of 42.78 k Da and the theoretical isoelectric point was of 6.57.The calculated instability parameter was of 35.92, which indicated that this protein was stable in classification.The secondary structure of resveratrol synthase mainly contained α-helix, random curling and β-sheet, which content was of 44.13%, 26.53% and 17.66%, respectively.