期刊论文

Wu, Y.-M.

中文

中华微生物学和免疫学杂志

0254-5101

2011

31

9

781-786

10.3760/cma.j.issn.0254-5101.2011.09.004

传染病防治重大专项课题项目（2008ZX10004-001）； 国家自然科学基金项目（81001318）；

本校为第一且通讯机构

目的 检测鹦鹉热嗜衣原体（Chlamydophila psittaci，Cps）蛋白酶样活性因子（chlamydial protease-like activity factor，CPAF）免疫优势区基因在E.coli BL21中高效表达的产物在Cps感染诊断中的应用，为建立Cps感染的快速诊断奠定实验基础。 方法 利用生物信息学软件筛选出Cps CPAF免疫优势区基因序列（CPAFm， A196～A450）,设计特异性引物, PCR扩增目的基因, 将其克隆入pGEX6p-2载体后转化E.coli BL21,利用IPTG诱导表达重组蛋白，Western blot鉴定重组蛋白。以纯化的重组蛋白作为包被抗原，建立血清学诊断的间接ELISA法；同时用商品ELISA试剂盒与建立的间接ELISA 法检测180份可疑Cps感染的病...

Objective: To clone and express the immunodominant domain of the chlamydial protease-like activity factor(CPAF) from Chlamydophila psittaci(Cps) and evaluated the diagnosing value of the recombinant protein in Cps infection. Methods: The immunodominant region epitope of CPAF (CPAFm, A196-A450) from Cps was chosen according to bioinformatics analysis and references. The specific primer was designed and the gene was amplified by PCR and then ligated into a pGEX6p-2 vector. Recombinant protein was induced to express by IPTG and analyzed by SDS-PAG...