摘要： 根据GenBank已发表的PGIP基因序列设计引物,通过PCR直接扩增的方法从青花菜 (Brassica oleracea var.italica P)中克隆了一个新的PGIP基因,命名为BoPGIP1.BoPGIP1基因序列全长为1 102 bp,由2个外显子和1个内含子构成,外显子总长为993 bp,编码330个氨基酸序列,分子量为37 kDa.对推导的氨基酸序列进行分析,发现该序列包含个N-糖基化位点,2个蛋白激酶C磷酸化位点,个酪蛋白激酶Ⅱ磷酸化位点,4个N-豆蔻酰化位点.将BoPGIP1氨基酸序列与数据库中的其他植物PGIP进行同源序列比对,构建系统进化树,发现所比对基因都含有LRR结构,在14位点的LRR结构域在所有物种中极其保守;系统树显示,该序列最先与甘蓝型油菜和拟南芥聚类,其次和棉花、番茄和豆科作物的PGIP聚类,基本符合按形态特征分类的亲缘关系.

关键词: BoPGIP1, 克隆, 序列分析, 生物信息学

Abstract: BoPGIP1 gene was cloned from Brassica. oleraca var. italica P by PCR amplification,with two pairs of primers designed according to the sequences of other known pgip genes from GenBank. The full length of BoPGIP 1 gene is 1 102 bp and the largest open reading frame is 993 bp encodes a protein of 330 amino acids with a predicted molecular mass of 37 kDa,interrupted by one intron of 95 bp in length. Sequence analysis revealed that it has five potential N2glyco2 sylation sites,two protein kinase C phosphrylation sites,five casin kinase II phosphrylation sites,four N2myristoylation sites. The amino acids sequences alignment confirmed that 145 LRR stucture was highly conserved in all aligned PGIP se2 quences. The results of phylogenetic analysis showed that BoPGIP1 and PGIP from Brassica napus and Arabidopsis thaliana clustered together firstly,and then came from Gossypium barbadense,Lycopersicon esculentum and Phaseolus vulgaris in turn. The clustering results basically accorded with morphological classification system.

Key words: BoPGIP1,Cloning, Sequence analysis, Bioinformatic research

中图分类号: 

• Q78