CAJ | 학술논문

本研究通过建立CSFV噬菌体展示多肽库,并对其进行生物淘选,以期获得E2蛋白上新的抗原表位.选择CSFV石门株(SM)和疫苗株(HCLV)为代表株,采用噬菌体展示技术,以T7select415-1b为载体,分别构建了CSFV SM株和HCLV株E2基因噬菌体展示多肽库(SM-E2库和HCLV-E2库).通过生物淘选和噬菌体原位杂交技术,采用7株猪瘟单克隆抗体和1株猪瘟高免血清分别对构建的SM-E2库和HCLV-E2库进行抗原表位筛选.结果共筛选到了5条与E2蛋白高度同源的序列,在E2蛋白上的同源区域分别为TAVSPTTLR、YYEP、TTWKEYSH、GGQ(V)VK和PDGLPHY.结果表明,TAVSPTTLR、YYEP和TTWKEYSH序列与目前已知E2蛋白表位一致,说明它们是E2蛋白上的优势表位;GGQ(V)VK和PDGLPHY序列与预测表位一致,推测是E2蛋白上潜在的抗原表位.
본연구통과건립CSFV서균체전시다태고,병대기진행생물도선,이기획득E2단백상신적항원표위.선택CSFV석문주(SM)화역묘주(HCLV)위대표주,채용서균체전시기술,이T7select415-1b위재체,분별구건료CSFV SM주화HCLV주E2기인서균체전시다태고(SM-E2고화HCLV-E2고).통과생물도선화서균체원위잡교기술,채용7주저온단극륭항체화1주저온고면혈청분별대구건적SM-E2고화HCLV-E2고진행항원표위사선.결과공사선도료5조여E2단백고도동원적서렬,재E2단백상적동원구역분별위TAVSPTTLR、YYEP、TTWKEYSH、GGQ(V)VK화PDGLPHY.결과표명,TAVSPTTLR、YYEP화TTWKEYSH서렬여목전이지E2단백표위일치,설명타문시E2단백상적우세표위;GGQ(V)VK화PDGLPHY서렬여예측표위일치,추측시E2단백상잠재적항원표위.
The phage display peptides libraries of Classic Swine fever virus (CSFV)E2 gene were constructed to identify the potential epitopes of E2 protein by biopanning. The E2 genes of Shi-men strain (SM) and hog cholera lapinized virus (HCLV) strain were cloned into T7seclect-415b vector respectively. So the E2 gene phage display peptides libraries of Shimen strain and HCLV strain were constructed and named as SM-E2 libraries and HCLV-E2 libraries. CSFV polyclonal antibody(PcAB) and seven strains of McAbs were used to screen the epitopes in SM-E2 libraries and HCLV-E2 libraries by biopanning and phage in situ hybridization. After analyzing the se-quences of selected phage recombinants, five sequences which were highly homologous with TAVSPTTLR, YYEP, TTWKEYSH, GGQ(V)VK and PDGLPHY were selected by the MAbs and CSFV PcAB. These results proved that these three sequences of TAVSPTTLR, YYEP and TTWKEYSH are dominant epitopes on E2 protein, which were reported in the previous papers. In addition, two sequences of GGQ(V)VK and PDGLPHY corresponds to the epitopes predicted by computer. So these two sequences might be the potential epitopes of E2 protein.