【摘要】 目的利用噬菌体12肽库筛选能与EB病毒(EBV)LMP1 CTAR23蛋白高效结合的短肽。方法利用亲和层析柱加酶切的方法纯化CTAR23蛋白,并用其筛选噬菌体12肽库。通过夹心ELISA法分析噬菌体克隆与CTAR23蛋白的亲和力。通过硝酸纤维膜斑点印迹法进行阳性克隆鉴定,并对阳性克隆进行测序及序列分析。结果对12肽库进行3轮筛选后,噬菌体克隆具有良好的富集效果。ELISA提示筛选出来的噬菌体克隆能与CTAR23蛋白高效结合。硝酸纤维膜斑点印迹法证实阳性率达到100%,测序结果显示噬菌体上的短肽有共同序列HVSLHKHYPTAS。结论噬菌体短肽HVSLHKHYPTAS为研究LMP1致瘤机理、开发拮抗LMP1蛋白的小分子短肽类药物奠定坚实的基础。更多还原

【Abstract】 Objective Use phage display peptide library to identify and characterize the peptide that is able to bind with epstein-barr virus(EBV) LMP1 CTAR23.Methods Affinity chromatography and enzyme action on column were used to purify CTAR23 protein,by which a random phage display peptide library was screened.The clones were identified by sandwich ELISA,dot blotting and DNA sequencing.Results All the selected clones showed positive signal in sandwich ELISA and dot blotting.The amino acid sequences deduced from DNA sequences shared the same sequence as in phage display clone:HVSLHKHYPTAS.Conclusions Our results suggested that the peptide HVSLHKHYPTAS could bind with EBV LMP1 CTAR23.It provided a basis for further research on the oncogenic role of LMP1,the gene therapy and vaccines for NPC.更多还原