论文部分内容阅读
目的探讨是否能够通过噬菌体展示技术筛选与肠致病性大肠埃希菌(EPEC)关键蛋白EspB结合的短肽。方法采用体外大肠埃希菌中表达和纯化EspB蛋白情况,Western blot验证蛋白的表达;Ph.D.12-蛋氨酸噬菌体展示技术用来筛选EspB特异结合蛋白;ELISA方法用来鉴定这些特异性结合蛋白与EspB的亲和力。结果 Western blot验证从pET21b-EspB转染质粒中提取的EspB蛋白。噬菌体展示技术筛选出了噬菌体6、7、8、12候选蛋白,ELISA检测结果显示这些候选蛋白与EspB蛋白的亲和力高于对照蛋白。结论我们的数据提供了一种潜在的研究策略即通过靶向结合EspB蛋白可抑制EPEC对上皮细胞的粘附。
Objective To explore whether phage display technology can be used to screen short peptides bound to EspB, a key protein of enteropathogenic Escherichia coli (EPEC). Methods The EspB protein was expressed and purified in Escherichia coli in vitro and the protein expression was confirmed by Western blot. Ph.D.12-methionine phage display technique was used to screen EspB specific binding protein. The ELISA method was used to identify these specific binding proteins Affinity with EspB. Results Western blot confirmed the EspB protein extracted from plasmid pET21b-EspB. Phage display technology was selected phage 6,7,8,12 candidate protein, ELISA test results showed that these candidate proteins and EspB protein affinity higher than the control protein. Conclusions Our data provide a potential research strategy to inhibit the adhesion of EPEC to epithelial cells by targeting the binding of EspB protein.