根据GeneBank中公布的GGPPS基因保守区设计特异引物,克隆南方红豆杉GGPPS基因的DNA和cDNA序列,并通过生物信息学方法对其核苷酸序列和蛋白质结构进行分析预测。结果表明,GGPPS基因DNA和cDNA序列都为1 182 bp,DNA序列中无内含子,cDNA序列为一个编码393个氨基酸残基的开放式阅读框;GGPPS的理论相对分子质量为42 590,等电点为5.68。核苷酸同源性分析显示,它与Taxus canadensis(AF081514)同源性为98.8%;推导出的氨基酸序列与Taxus canadensis(AAD16018)同源性达99.0%。利用Protparam、SignalP、ProtScale、SOPMA、Swiss-Modeling、Scan Prosite和MEGA4.0等生物信息学工具分别对其理化性质、信号肽、疏水性、亲水性、二级结构、三级结构和进化树进行分析,为其功能研究和生物合成紫杉醇研究提供分子基础。
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In this study,the DNA sequence and cDNA sequence of ggpps were cloned from Taxus wallichiana var.Mairei.Results showed that the length of both DNA and cDNA sequences is 1182 bp,without introns,contained an opening reading frame of 1182 bp,which coded for 393 amino acid residues;theoretical molecular weight is 42.59kDa,and isoelectric point is 5.68.Alignment showed that cDNA sequence exhibited 98.8 % similarity to the ggpps gene of Taxus canadensis(AF081514) which has been reported in NCBI,and its protein sequence has 99.0 % similarity to GGPPS of Taxus canadensis(AAD16018).The bioinformatics of GGPPS,including physical and chemical properties,the signal peptide,hydrophobicity,hydrophilicity,secondary structure,tertiary structure,and phylogenetic trees,were analyzed with bioinformatics softwares and database,such as Protparam,SignalP,ProtScale,SOPMA,Swiss-Modeling,Scan Prosite,MEGA4.0 and so on.
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