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以结球甘蓝(Brassica oleracea L.var.capitata L.)E1花粉为试材,对萌发前后的花粉总蛋白进行双向电泳及差异蛋白质谱分析,发现延伸因子BoEF-Tu蛋白在花粉萌发后较萌发前表达上调。利用同源克隆得到甘蓝BoEF-Tu基因的全长cDNA序列,其长度为1 728 bp,具有一个完整开放阅读框(ORF),位于112～1 473 bp处,编码453个氨基酸残基,预测分子量为49.17 kD,等电点为6.52。生物信息学分析表明:BoEF-Tu蛋白含有9个α–螺旋结构,18个β–折叠结构,不含信号肽和跨膜区,在106～121位氨基酸残基处有1个GTP结合区域(DKAPEEKKRGITIATA)。氨基酸同源性分析表明,BoEF-Tu与拟南芥EF-TuM同源性较高,达到93%;系统进化分析表明,BoEF-Tu与拟南芥EF-Tu的亲缘关系最近。
The pollen total protein before and after germination was analyzed by two-dimensional gel electrophoresis and differential protein profile analysis using the E1 pollen of Brassica oleracea L.var.capitata L. The results showed that the elongation factor BoEF-Tu protein was more germinated after pollen germination Pre-expression increased. The full-length cDNA sequence of BoEF-Tu was obtained by homologous cloning. It has a length of 1 728 bp and a complete open reading frame (ORF) located at 112-1473 bp encoding 453 amino acid residues. The predicted molecular weight 49.17 kD, isoelectric point of 6.52. Bioinformatics analysis showed that the BoEF-Tu protein contains 9 α-helical structures, 18 β-sheet structures, no signal peptide and transmembrane region, and one GTP binding region at the amino acid residues 106-121 DKAPEEKKRGITIATA). Amino acid homology analysis showed that BoEF-Tu had high homology with EF-TuM of Arabidopsis thaliana, reaching 93%. Phylogenetic analysis showed that BoEF-Tu had the closest genetic relationship with Arabidopsis EF-Tu.