甘蓝型油菜防御素基因的克隆与生物信息学分析

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RT-PCR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cDNA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。

Cloning and Bioinformatics Analysis of Defensin Gene from Brassica napus

The aims were to investigate the structure and function of Brassica rapa defensin genes. According to the sequence of Brassica rapa defensin genes, 4 Brassica napus defensin genes were cloned by polymerase chain reaction (PCR) and predicted by bioinformatic methods. The results showed that the cDNA sequence was 325-335 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 tree showed 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, which might be secreted protein.