期刊论文

Wu, Y.

中文

中华微生物学和免疫学杂志

0254-5101

2014

34

8

604-608

10.3760/cma.j.issn.0254-5101.2014.08.007

81202323:国家自然科学基金 31270218:国家自然科学基金 81273026:国家自然科学基金 2012FJ6009:湖南省科技厅项目 B2012-042:湖南省卫生厅项目

本校为第一且通讯机构

目的：构建鹦鹉热嗜衣原体（ Chlamydophila psittaci，Cps） CPSIT_p7原核表达载体，表达并纯化CPSIT_p7，检测其免疫原性，观察其在持续性感染过程中的mRNA和蛋白动态表达情况。方法原核表达和纯化His-CPSIT_p7融合蛋白，免疫BALB/c小鼠制备多克隆抗体，ELISA分析His-CPSIT_p7蛋白的免疫原性。采用青霉素与Cps共同处理细胞来制备持续性感染模型， RT-PCR和Western blot检测CPSIT_p7在其持续性感染过程中的表达情况。结果成功构建了pET30a-CPSIT_p7原核表达载体，并表达和纯化较稳定的重组蛋白；该蛋白免疫小鼠40 d后，ELISA 检测血清特异性抗体效价为1∶1000000；RT-PCR和Western blot 结果显...

Objective: To construct a prokaryotic expression plasmid for CPSIT_p7 gene from Chlamydophila psittaci (Cps) 6BC strain and to evaluate immunogenicity of the recombinant protein His-CPSIT_p7 and detect its dynamic expression at mRNA and protein levels in HeLa cells during persistent Cps infection.Methods: The fusion protein His-CPSIT_p7 was expressed in E. coli BL21 and purified by Ni-NTA affinity chromatography. BALB/c mice were immunized with the recombinant protein to prepare polyclonal antibody for evaluation of the immunogenicity of His-CP...