禽巴氏杆菌株粘附蛋白基因的克隆和序列分析

用PCR从禽巴氏杆菌P-1059基因组DNA中分别扩增编码粘附蛋白OmpH,PtfA,PlpE和PM1665的基因序列,将PCR产物克隆到pMD18-T载体中,转化大肠杆菌DH5α,经菌落PCR筛选含有重组质粒DNA的重组子,并对目的基因进行测序.DNA测序结果表明,P-1059株ompH,plpE,ptfA和pm1665基因大小分别为1 032,1 008,435,348bp,分别编码343,335,144,115个氨基酸的多肽.Blast分析结果显示,P-1059株与已报道不同血清型巴氏杆菌ompH基因的同源性为95%~97%,与ptfA基因的同源性为95%~99%,与plpE基因的同源性为94%~97%,而与pm1665基因的同源性为99%.克隆得到的ompH,ptfA,plpE和pm1665基因,为禽巴氏杆菌粘附因子及其致病机理的研究奠定基础.

Cloning and Sequencing of Adhesive Protein Genes of Avian Pasteurella Multocida

The genes encoding the outer membrane protein H (OmpH),lipoprotein E (PlpE),type 4 fimbrial subunit protein (PtfA) and fibronectin binding protein (PM1665) were amplified by PCR from genomic DNA of avian Pasteurella multocida P-1059,respectively.The PCR products were cloned into the pMD18-T vector and then sequenced.Sequence analyses showed that the open reading frames (ORFs) of the ompH,plpE,ptfA and pm1665 genes were 1 032,1 008,435 and 348 bp in length,and encoded the 343,335,144 and 115 amino acid residues.Nucleotide sequence homologies of the ompH,plpE,ptfA and pm1665 genes between the P-1059 and the previously reported different serotype strains of P. multocida were 95%~97%,95%~99%,94%~97% and 99%,respectively.In conclusion,the cloned genes encoding the adhesive proteins of avian P. multocida P-1059 will contribute to further study on pathogenesis of this organism.