Mullen Lisa M, Nair Sean P, Ward John M, Rycroft Andrew N, Williams Rachel J, Henderson Brian
Division of Microbial Diseases, UCL Eastman Dental Institute, University College London, London WC1X 8LD, United Kingdom.
Vet Microbiol. 2007 May 16;122(1-2):123-34. doi: 10.1016/j.vetmic.2006.12.022. Epub 2007 Jan 26.
The Pasteurellaceae contain a number of important animal pathogens. Although related, the various members of this family cause a diversity of pathology in a wide variety of organ systems. Adhesion is an important virulence factor in bacterial infections. Surprisingly little is known about the adhesins of the Pasteurellaceae. To attempt to identify the genes coding for adhesins to some key components of the hosts extracellular matrix molecules, phage display libraries of fragmented genomic DNA from Haemophilus influenzae, Actinobacillus pleuropneumoniae, Pasteurella multocida and Aggregatibacter actinomycetemcomitans, were prepared in the phage display vector pG8SAET. The libraries were screened against human or porcine fibronectin, serum albumin or a commercial extracellular matrix containing type IV collagen, laminin and heparin sulphate. Four genes encoding putative adhesins were identified. These genes code for: (i) a 34 kDa human serum albumin binding protein from Haemophilus influenzae; (ii) a 12.8 kDa fibronectin-binding protein from Pasteurella multocida; (iii) a 13.7 kDa fibronectin-binding protein from A. actinomycetemcomitans; (iv) a 9.5 kDa serum albumin-binding protein from A. pleuropneumoniae. None of these genes have previously been proposed to code for adhesins. The applications of phage display with whole bacterial genomes to identify genes encoding novel adhesins in this family of bacteria are discussed.
巴斯德氏菌科包含许多重要的动物病原体。尽管该科的各个成员具有相关性，但它们会在多种器官系统中引发多种病理变化。黏附是细菌感染中的一个重要毒力因子。令人惊讶的是，人们对巴斯德氏菌科的黏附素知之甚少。为了尝试鉴定编码针对宿主细胞外基质分子某些关键成分的黏附素的基因，使用噬菌体展示载体pG8SAET构建了来自流感嗜血杆菌、胸膜肺炎放线杆菌、多杀性巴氏杆菌和伴放线聚集杆菌的基因组DNA片段的噬菌体展示文库。这些文库针对人或猪纤连蛋白、血清白蛋白或含有IV型胶原、层粘连蛋白和硫酸乙酰肝素的商业细胞外基质进行筛选。鉴定出了四个编码假定黏附素的基因。这些基因编码：（i）来自流感嗜血杆菌的一种34 kDa人血清白蛋白结合蛋白；（ii）来自多杀性巴氏杆菌的一种12.8 kDa纤连蛋白结合蛋白；（iii）来自伴放线聚集杆菌的一种13.7 kDa纤连蛋白结合蛋白；（iv）来自胸膜肺炎放线杆菌的一种9.5 kDa血清白蛋白结合蛋白。此前没有提出过这些基因编码黏附素。本文讨论了利用噬菌体展示全细菌基因组来鉴定该细菌家族中编码新型黏附素的基因的应用。
