本试验以芥蓝为试验材料,通过同源序列检索分析设计特异性引物,采用反转录PCR方法成功分离出芥蓝GGPPS1基因.BoaGGPPS1基因CDS全长为1113 bp,编码370个氨基酸.生物信息学分析结果显示,BoaGG-PPS1蛋白的等电点为6.07,相对分子量为39790,不含跨膜结构域和信号肽.序列分析结果显示,BoaGGPPS1与其他植物GGPPS高度保守,与结球甘蓝的一致性最高,达到98%.系统进化树分析结果表明,BoaGGPPS1与十字花科其他植物的亲缘关系较近,与结球甘蓝的亲缘关系最近.BoaGGPPS1基因在芥蓝不同器官中均有表达,其中叶片中的表达量最高,根系中的表达量最低.构建原核表达载体pEASY-Blunt E1-BoaGGPPS1,并对其进行诱导表达,结果发现该蛋白在大肠杆菌体内主要以包涵体的形式存在.%Using Chinese kale (Brassica oleracea var. alboglabra) as experimental material, the BoaGGPPS1 gene encoding geranylgeranyl pyrophosphate synthase (GGPPS1) was successfully isolated by reverse transcription PCR and the specific primers were designed by homologous sequence analysis. The coding sequence length of BoaGGPPS1 gene was 1 113 bp,encoding 370 amino acids. Bioinformatics analysis results showed that the isoelectric point of BoaGGPPS1 protein was 6.07,and the relative molecular weight was 39 790,BoaGGPPS1 protein contained no transmembrane domain and sig-nal peptide. The sequence analysis results showed that BoaGGPPS1 was highly conserved and had high identity with cab-bage,reached 98%. Phylogenetic tree analysis results showed that BoaGGPPS1 was more closely related to other plants of Brassicaceae, and had the closest relationship with Brassica oleracea var. capitata. The expression of BoaGGPPS1 gene was found among different organs of Chi-nese kale, and the highest expression level was found in leaves,whereas the lowest expression was found in roots. The prokaryotic expression vector pEASY-Blunt E1-BoaGG-PPS1 was constructed, and the protein was expressed in the form of inclusion bodies in Escherichia coli.
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