Abstract Abstract Porcine contagious pleuropneumonia (PCP) is a respiratory disease caused by Actinobacillus pleuropneumoniae (APP) infection, which is one of the 5 major epidemics in the world's pig industry. Based on APP OmpP2 sequences in GenBank, the primers were designed. APP OmpP2 was amplified by PCR from APP genome, sequenced and submitted to NCBI (Access No. KM357903). Bioinformatics analysis showed that the APP OmpP2 gene was composed of 1 131 bp, encoding a 376 amino acid protein. This protein was a porin protein with high homology among different APP serotypes. APP OmpP2 gene was subcloned into pET32a (+) to construct recombinant plasmid pET-OmpP2. rOmpP2 expression in Escherichia coli BL21 was induced by isopropyl β-D-1- thiogalactopyranoside (IPTG), and determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) and Western blot. The results showed that rOmpP2 with 42 kD was detected. Animal experiment was conducted by BALB/c mice (Mus musculus). The serum immunoglobulin G (IgG) level was detected by enzyme-linked immuno sorbent assay (ELISA). The results showed that survival rate of infected mice with aluminium adjuvant+50 μg rOmpP2, aluminium adjuvant+30 μg rOmpP2 and aluminium adjuvant+10 μg rOmpP2 were 50%, 30% and 10%, respectively, and all mice died in adjuvant group and control group. The surviving mice slowly returned to normal health status. Mice that were subcutaneously immunized with rOmpP2 showed an partial protection against the challenge with APP. This study provides basic data for screening new molecular vaccines of porcine contagious pleuropneumonia.
HU Pei-Pei,YANG Ting-Ting,HUANG Bi-Shen, et al. Immune Protection Research Against Challenge Infection in Mice (Mus musculus) Induced by APP rOmpP2 of Porcine (Sus scrofa)[J]. , 2017, 25(2): 299-306.
[1]刘 霞，刁有祥，禚宝山.猪传染性胸膜肺炎的研究进展[J].山东农业大学学报(自然科学版), 2006, 37 (1): 152~156 [2]韩国全，郭万柱，王利娜，李宇，李云云，王印，王小玉.猪传染性胸膜肺炎新型疫苗的研究进展[J].猪业科学,2009, 26(2):25-30. [3]杜爱庆，刁有祥，张伟，张瑞平，臧大鹏，刘芳娜.猪胸膜肺炎放线杆菌溶血毒素的克隆表达及其对小鼠免疫保护作用[J].微生物学报,2008,48(3):342~348. [4]鱼艳荣，刘希成.革兰氏阴性菌外膜蛋白研究进展[J].动物医学进展,2000,21(2):35-39. [5]赵香汝，杨汉春.细菌外膜蛋白的研究现状[J].中国兽医杂志，1997，23(12):41-42. [6]Xuehe Hu, Hao Yan, Ke Liu, Jiansheng Hu, Chao Qi, Jihong Yang, Yanli Liu, Jin Zhao and Jinlin Liu.Identification and characterization of a novel stress-responsive outer membrane protein Lip40 from Actinobacillus pleuropneumoniae[J]. BMC Biotechnology，2015，15:106-127. [7]Shao M, Wang Y, Wang C, Guo Y, Peng Y, Liu J, Li G, Liu H, Liu S.Evaluation of multicomponent recombinant vaccines against Actinobacillus pleuropneumoniae in mice[J]. Acta Vet Scand. 2010;52:52. [8]Van den Bosch H, Frey J.Interference of outer membrane protein PalA with protective immunity against Actinobacillus pleuropneumoniae infections in vaccinated pigs[J]. Vaccine. 2003，21(25-26):3601-3607. [9]Del Pozo Sacristán R1, Michiels A，Martens M，Haesebrouck F，Maes D.Efficacy of vaccination against Actinobacillus pleuropneumoniae in two Belgian farrow-to-finish pig herds with a history of chronic pleurisy[J].Vet Rec. 2014,174(12):302. [10]Wang C，Wang Y，Shao M，Si W，Liu H，Chang Y，Peng W，Kong X，Liu S.Positive role for rApxIVN in the immune protection of pigs against infection by Actinobacillus pleuropneumoniae[J].Vaccine.2009,27(42):5816-5821. [11]彭秀玲,袁汉英,谢毅.基因工程实验技术[M].长沙:湖南科技出版社,1998. 183-188. [12]刘建杰,陈焕春,何启盖,吴 斌,李 冲,徐晓娟,陈汉阳.猪传染性胸膜肺炎新型亚单位菌苗对小鼠的效力研究[J].畜牧兽医学报,2005,36(2):177-180
