摘要
本研究将鹦鹉热衣原体的MOMP基因经PCR扩增后与pET-22b(+)载体连接构建重组质粒,转化入E.coli,通过IPTG诱导表达MOMP蛋白,鉴定重组蛋白的表达和反应性。结果显示,重组蛋白被成功表达,分子质量大小为40 ku,经溶解、透析纯化后,重组MOMP蛋白纯度为91%,能与鼠抗重组MOMP蛋白抗体产生特异性反应,与鹦鹉热衣原体阳性血清呈强阳性反应。重组MOMP蛋白与ISA201佐剂乳化制成鹦鹉热衣原体亚单位疫苗,免疫绵羊后抗体效价高达1∶4222,用强毒株攻毒后保护率达100%,表明此鹦鹉热衣原体亚单位疫苗具有良好的免疫原性和免疫保护性。
The MOMP gene of Chlamydia psittaci was amplified by PCR and cloned into the p ET-22 b(+)expression vector to construct the recombinant plasmid.The recombinant plasmid was transformed into E.coli and induced by IPTG,then the recombinant protein was identified.Results showed that the recombinant protein was successfully expressed with a molecular weight of 40 ku.It’s purity was 91%after dialysis and purification.The recombinant MOMP could specific react with mouse antibody against the MOMP protein and exhibit strongly positive with the positive serum of Chlamydia psittaci.The recombinant MOMP protein mixed with ISA201 adjuvant to prepare the subunit vaccine of Chlamydia psittaci.The vaccine immunized the sheep and the antibody titers was as high as 1∶4222.After the wild-type strain challenge,the protection rate of this vaccine was 100%.All these results demonstrated that the subunit vaccine of Chlamydia psittaci could induce robust immunogenicity and protective efficacy.
作者
张静
李敏
刘少蓉
高强
张伟
杨茹
董杰
段跃强
ZHANG Jing;LI Min;LIU Shao-rong;GAO Qiang;ZHANG Wei;YANG Ru;DONG Jie;DU AN Yue-qiang(Inner Mongolia Huaxi Biological Technology Co.,Ltd.,Huhhot 010111,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2020年第7期869-873,共5页
Chinese Veterinary Science
基金
呼和浩特市科技计划项目(2018-高-重-4)。
