中国免疫学杂志2018，Vol.34Issue(5)：653-657,5.DOI:10.3969/j.issn.1000-484X.2018.05.003
从T7噬菌体展示cDNA文库筛选生殖支原体黏附蛋白的互作蛋白
Identification of protein that interacts with adhesion protein of Mycoplasma genitalium from T7-phage display cDNA library
摘要
Abstract
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 My-coplasma 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.According to the results of DNA sequencing and BLAST analysis,the selected randomly 32 positive clones included 7 kinds different sequences,of which the number of RPL35 repeats was the most.The results of indirect ELISA,Dot immunoblot and Far-western blot showed that 7 representative phages could bind specifically with rMgPa.Conclusion:60S ribosomal protein L35(RPL35) may be the interacting protein of MgPa,which lays the experimental foundation for understanding the function of MgPa and the pathogenesis of Mycoplasma genitalium.关键词
T7噬菌体/生殖支原体/MgPa/互作蛋白Key words
T7 phage/Mycoplasma genitalium/MgPa/Interacting proteins分类
医药卫生引用本文复制引用
戴佩,邓湘赢,余敏君,李玲玲,罗丹,龙娴,曾焱华..从T7噬菌体展示cDNA文库筛选生殖支原体黏附蛋白的互作蛋白[J].中国免疫学杂志,2018,34(5):653-657,5.基金项目
国家自然科学基金项目(No.31370207)和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. （No.31370207）
