Clinical Bacteriology Laboratory, Indian Veterinary Research Institute (IVRI), Mukteswar 263138, Nainital (District), Uttarakhand (UK), India.
Microb Pathog. 2013 Dec;65:48-56. doi: 10.1016/j.micpath.2013.09.007. Epub 2013 Oct 9.
Pasteurella multocida serogroup B:2, a causative agent of haemorrhagic septicaemia (HS) in cattle and buffalo especially in tropical regions of Asian and African countries, is known to possess several outer membrane proteins (OMPs) as immunogenic antigens. In the present study, omp87 gene encoding for 87 kDa OMP (Omp87) protein of P. multocida serogroup B:2 strain P52, has been amplified (∼2304 bp), cloned in to pET32a vector and over-expressed in recombinant Escherichia coli as fusion protein. The recombinant Omp87 protein (∼102 kDa) including N-terminus hexa-histidine tag was purified under denaturing condition. Immunization of mice with rOmp87 resulted in increased antigen specific IgG titres in serum and provided protection of 66.6 and 83.3% following homologous (B:2) and heterologous (A:1) challenge, respectively. A homology model of Omp87 revealed the presence of two distinct domains; N-terminal domain with four POTRA repeats in the periplasmic space and a pore forming C-terminal β-barrel domain (β1- β16) in the outer membrane of P. multocida, which belong to Omp85-TpsB transporter superfamily of OMPs. The study indicated the potential possibilities to use rOmp87 protein along with suitable adjuvant in developing subunit vaccine for haemorrhagic septicaemia and pasteurellosis in livestock.
多杀巴斯德菌血清型 B:2 是一种引起牛和水牛出血性败血症（HS）的病原体，尤其在亚洲和非洲国家的热带地区较为常见，该菌已知含有几种外膜蛋白（OMPs）作为免疫原性抗原。在本研究中，扩增了编码多杀巴斯德菌血清型 B:2 菌株 P52 的 87 kDa OMP（Omp87）蛋白的 omp87 基因（约 2304 bp），将其克隆到 pET32a 载体中，并在重组大肠杆菌中作为融合蛋白过表达。重组 Omp87 蛋白（约 102 kDa）包含 N 端六组组氨酸标签，在变性条件下进行纯化。用 rOmp87 免疫小鼠可增加血清中的抗原特异性 IgG 滴度，并在同源（B:2）和异源（A:1）攻毒后分别提供 66.6%和 83.3%的保护率。Omp87 的同源模型显示存在两个不同的结构域；N 端结构域在周质空间中含有四个 POTRA 重复序列，而在多杀巴斯德菌的外膜中存在一个形成孔的 C 端β-桶结构域（β1-β16），属于 OMPs 的 Omp85-TpsB 转运体超家族。该研究表明，有可能使用 rOmp87 蛋白与合适的佐剂一起开发用于家畜出血性败血症和巴氏杆菌病的亚单位疫苗。
