Singh Ajay Pratap, Singh Satparkash, Ranjan Rajeev, Gupta Santosh Kumar, Singh Vijendra Pal, Sharma Bhaskar
Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Bareilly- 243122, UP, India.
J Vet Sci. 2010 Sep;11(3):227-33. doi: 10.4142/jvs.2010.11.3.227.
Outer membrane proteins of Pasteurella (P.) multocida have been known to be protective immunogens. Pasteurella lipoprotein E (PlpE) has been reported to be an important cross reactive outer membrane protein in P. multocida. The gene encoding the PlpE of P. multocida serotypes A: 3, B: 2 and D: 1 was amplified from the genomic DNA. The amplified products were cloned and the nucleotide sequence was determined. Sequence analysis of the recombinant clones revealed a single open reading frame of 1,011 bp, 1,008 bp and 1,017 bp encoding a protein with a calculated molecular mass of 37.829 kDa, 37.389 kDa and 37.965 kDa for serotypes A: 3, B: 2 and D: 1 respectively. The comparison of the plpE sequence in different capsular types revealed a high degree (> 90%) of homology. Furthermore, the plpE gene of Haemorhhagic septicaemia causing serotype (B: 2) was expressed in E. coli and recombinant PlpE was strongly immunostained by antiserum against whole cell antigen, indicating that the protein is expressed in vivo.
多杀性巴氏杆菌(P. multocida)的外膜蛋白已知是保护性免疫原。据报道,多杀性巴氏杆菌脂蛋白E(PlpE)是一种重要的交叉反应性外膜蛋白。从血清型A:3、B:2和D:1的多杀性巴氏杆菌基因组DNA中扩增出编码PlpE的基因。对扩增产物进行克隆并测定核苷酸序列。重组克隆的序列分析显示,血清型A:3、B:2和D:1分别有一个1011 bp、1008 bp和1017 bp的单一开放阅读框,编码的蛋白质计算分子量分别为37.829 kDa、37.389 kDa和37.965 kDa。不同荚膜类型中plpE序列的比较显示出高度(>90%)同源性。此外,引起出血性败血症的血清型(B:2)的plpE基因在大肠杆菌中表达,重组PlpE被抗全细胞抗原血清强烈免疫染色,表明该蛋白在体内表达。