期刊论文

Wang, S.-P.

中文

中华微生物学和免疫学杂志

0254-5101

2012

32

8

706-710

10.3760/cma.j.issn.0254-5101.2012.08.008

国家自然科学基金（81072418） 湖南省教育厅优秀青年基金（098088） 湖南省高校科技创新团队资助项目（湘教通[2010]212号）

本校为其他机构

目的 了解肺炎支原体(Mycoplasma pneumoniae,Mp)P1蛋白第1125 ～1395氨基酸片段(P1C蛋白)的免疫学活性及其细胞黏附作用.方法 构建用于表达重组P1C片段(rP1C)的原核表达载体pGEX6p-2/p1c,采用SDS-PAGE和Western blot鉴定rP1C.采用基于GST的亲和层析法提纯rP1C,提纯的rP1C免疫BALB/c小鼠,ELISA检测小鼠抗rP1C血清的效价.采用Western blot检测rP1C对Mp感染患者血清的免疫反应性.采用间接免疫荧光法检测rP1C黏附HeLa细胞及其免疫血清黏附抑制作用.结果 所构建的原核表达系统能有效表达相对分子质量约为66×103的可溶性rP1C.rP1C免疫小鼠后,其抗血清ELISA效价高达1∶64000.rP1C能被Mp感染者血清及小...

Objective: To determine the immunogenic and adhesive abilities of a segment (P1C protein) that located at the carboxy terminal region of P 1 protein (1125 to 1395 amino acids). Methods: A recombinant prokaryotic vector (pGEX6p-2/p1c) was constructed for P1C protein expression in E. coli BL21DE3. The expressed target recombinant protein (rP1C) was identified using SDS-PAGE and Western blot assay, and then extracted by GST-based affinity chromatography. The purified rP1C was used to immunize BALB/c mice to obtain rP1C-antiserum and titer of the a...