甘蓝型油菜防御素基因的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of Defensin Gene from Brassica napus

为了研究甘蓝型油菜防御素基因的结构和功能,根据白菜防御素基因序列设计引物,采用RTPCR方法从甘蓝型油菜中克隆到4个防御素基因,并对其序列进行了生物信息学分析。结果表明,甘蓝型油菜防御素基因c DNA全长325~335 bp,包含231~243 bp的开放阅读框,编码76~80个氨基酸;防御素基因在长期进化过程中产生了显著的差异,但防御素含有的8个保守半胱氨酸残基均稳定存在;Bndef1、Bndef2、Bndef3与萝卜Rs-AFP1、诸葛菜Omdef亲缘关系较近,Bndef4与豇豆Vudef亲缘关系较近;4个防御素蛋白均为稳定蛋白,具有信号肽结构,可能为分泌蛋白。

The aims were to investigate the structure and function of Brassica rapa defensin genes. Accordingto the sequence of Brassica rapa defensin genes, 4 Brassica napus defensin genes were cloned by polymerasechain reaction（PCR） and predicted by bioinformatic methods. The results showed that the c DNA sequence was325-335 bp, and the open reading frame（ORF） was 231-243 bp which could encode a polypeptide of 78-80 amino acids. Brassica napus defensin genes resulted in significant differences in the long evolutionary process,resulting in significant differences, but the 6-8 conservative cysteine residues are stable. Phylogenetic treeshowed that Bndef1, Bndef2, Bndef3 had close genetic relationship with Raphanus sativus Rs- AFP1 and Orychophragmus violaceus Omdef, Bndef4 had close genetic relationship with Vigna unguiculata Vudef. The Brassica napus defensin genes defensin proteins were stable protein with a signal peptide structure, whichmight be secreted protein.