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采用RACE技术,从烟叶中克隆了一个新的编码GGPPS蛋白的基因,命名为Ntggpps3,并对其进行了生物信息学分析。Ntggpps3的cDNA序列为1251 bp,包含一个长度为1092 bp的开放阅读框,编码363个氨基酸。NtGGPPS3蛋白与其他植物GGPPSs高度保守。生物信息学预测结果表明,NtGGPPS3的分子量为39.5 kD,等电点为6.28,为亲水性蛋白,N端有叶绿体信号肽。二级结构主要由螺旋和无规则卷曲组成,三级结构含有两个富含天冬氨酸的区域,并通过同源建模构建了NtGGPPS3的3D模型。进化分析表明,植物的GGPPS分为大亚基和小亚基两个分支,而且NtGGPPS3属于大亚基。
Using RACE technique, a new gene encoding GGPPS protein was cloned from tobacco leaves and named as Ntggpps3, and bioinformatics analysis was carried out. The cDNA sequence of Ntggpps3 was 1251 bp and contained a 1092 bp open reading frame encoding 363 amino acids. NtGGPPS3 protein is highly conserved with other plant GGPPSs. Bioinformatics prediction showed that NtGGPPS3 had a molecular weight of 39.5 kD and an isoelectric point of 6.28, which was a hydrophilic protein with N-terminal chloroplast signal peptide. The secondary structure consists mainly of helix and random coil. The tertiary structure contains two aspartate-rich regions. The 3D model of NtGGPPS3 was constructed by homology modeling. Phylogenetic analysis showed that plant GGPPS is divided into two subunits: big subunit and small subunit, and NtGGPPS3 belongs to the large subunit.