Identification of protein that interacts with adhesion protein of Mycoplasma genitalium from T7-phage display cDNA library
10.3969/j.issn.1000-484X.2018.05.003
- VernacularTitle: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
- Author:
Pei DAI
1
;
Xiang-Ying DENG
;
Min-Jun YU
;
Ling-Ling LI
;
Dan LUO
;
Xian LONG
;
Yan-Hua ZENG
Author Information
1. 南华大学病原生物学研究所
- Keywords:
T7 phage;
Mycoplasma genitalium;
MgPa;
Interacting proteins
- From:
Chinese Journal of Immunology
2018;34(5):653-657
- CountryChina
- Language:Chinese
-
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.
