禽多杀性巴氏杆菌ompW基因的克隆与序列分析

提取禽多杀性巴氏杆菌分离株Pm27基因组DNA,参考GenBank中Pm70株ompW基因序列设计引物,应用PCR技术扩增了细菌ompW基因,将扩增片段克隆到pMD18-T载体后测序。生物信息学分析表明,该基因编码区长度为615bp,编码204个氨基酸;氨基酸序列与参考株Pm70 OmpW蛋白的同源性最高,达98%;与睡眠嗜血杆菌的同源性77%,与琥珀酸产生菌的同源性为69.3%,与霍乱弧菌、大肠杆菌、耶尔森菌的同源性为50%左右。OmpW蛋白前21个氨基酸残基为信号肽序列,潜在的糖基化位点位于第122位氨基酸残基。

Cloning and Sequencing of ompW Gene of Avian Pasteurella multocida

The genomic DNA of avian Pasteurella multocida Pm27 strain was extracted.According to the published ompW gene sequence of Pm70 in GenBank,the primers were designed.The ompW gene of Pm27 was amplified by PCR and then cloned into pMD18-T plasmids.Sequencing and analyzing by method of bioinformatics showed that ompW ORF included 615 base pairs,encoding 204 amino acids.It shared 98%,77% and 69.3% homologies in amino acid level with strain Pm70(avian Pasteurella multocida),Haemophilus somnus and Mannheimia succiniciproducens,respectively.The amino acid homology between Pm27 and Vibrio alginolyticus,Yersinia pestis and Escherichchia coli was about 50%.The initial 21 amino acids appeared to be a signal peptide.The N-glcosylation site in mature peptides located at 122 amino acid.