Cloning, expression and purification of Proteus mirabilis OmpA in Pichia pastoris

摘要：

　　To produce an effective vaccine to protect chickens against Proteus mirabilis infection, OmpA, one of the major outer membrane proteins of Proteus mirabilis, was expressed in cells of the Pichia pastoris.After induction for 96 h in the culture with 1.0% methanol, the concentration of the expressed protein reached 8.0 μtg/mL.Expression of the recombinant protein was confirmed by SDS-PAGE and Western blot analysis using antibody against Escherichia coli-expressed OmpA probe.Taishan Robinia pseudoacacia polysaccharide (TPPPS), a plant-derived adjuvant, was mixed with recombinant OmpA vaccine, and subcutaneously vaccinated chickens.ELISA analysis indicated that a significant antibody response against OmpA was elicited in the vaccinated group.A high (80.0%) immuno-protection against Proteus mirabilis was observed in the challenge experimental chickens.These results suggested that TPPPS could be used as a plant-derived adjuvant to enhance the immunogenicity of recombinant P mirabilis OmpA vaccine, and recombinant P.mirabilis OmpA that mixed with TPPPS was an effective vaccine to protect chickens against P mirabilis infection.

关键词： Expression Pichia pastoris Proteus mirabilis Recombinant subunit vaccine Robinia pseudoacacia polysaccharide Adjuvant

作者: Yongbing Zhang Liping Liu Xiaodong Jiang Mingxu Shao Shanshan Chi Chuanwen Wang Cuilian Yu Ruiliang Zhu

作者单位: Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Technology, Shandong Agricultural University, 61 Daizong Avenue, Taian, Shandong 271018, PR China

会议类型: 国内会议

会议名称: 世界中医药学会联合会中医药抗病毒研究专业委员会第一届学术年会

会议地点: 烟台

会议语种:英文

页码: 190-206

在线出版日期: 2015-07-01（万方平台首次上网日期，不代表论文的发表时间）