期刊论文

中文

中国免疫学杂志

1000-484X

2018

34

5

653-657

10.3969/j.issn.1000-484X.2018.05.003

国家自然科学基金项目（No.31370207） 湖南省大学生研究性学习和创新性实验计划（湘教通2015-269号）联合资助

本校为第一机构

目的:从人尿道上皮细胞( SV-HUC-1) T7噬菌体展示cDNA文库筛选生殖支原体黏附蛋白( MgPa)的互作蛋白.方法:以原核表达、纯化的重组生殖支原体黏附蛋白( rMgPa)为靶分子,对人尿道上皮细胞T7噬菌体展示cDNA文库进行4轮生物淘选,采用T7特异性引物扩增阳性克隆的插入片段,PCR产物测序后进行BLAST分析,间接ELISA、斑点免疫印迹和Far-western blot检测阳性噬菌体与rMgPa的特异结合.结果:经过4轮生物淘选,噬菌体克隆富集明显; DNA测序后经BLAST比对分析,结果表明随机挑取的32个阳性克隆包括7种不同序列,其中以RPL35重复次数最多;间接ELISA、斑点免疫印迹和Far-western blot结果表明7个代表性噬菌体均能...

Objective: To screen and identify the protein that interacts with the adhesion protein of Mycoplasma genitalium ( MgPa) from T7-phage display cDNA library of human uroepithelial cells( SV-HUC-1) . Methods: Recombinant adhesion protein of Mycoplasma genitalium( rMgPa) was used as target molecule to biopan the T7 phage display cDNA library of SV-HUC-1 cell, the selected positive clones were analysed using DNA sequencing and BLAST analysis and identified by means of indirect ELISA,Dot immunoblot and Far-western blot. Results: After four rounds of biopanning,positive phages were obviously enriched...