摘要
目的 确定汉滩病毒（ＨＴＮＶ）上可与宿主细胞受体结合的一段配基表位。方法 型特异性ｍＡｂ ３Ｇ１，与 ＨＴＮＶ糖蛋白ＧＺ及核蛋白（ＮＰ）具有双结合活性，且中和效价很高。以纯化的 ｍＡｂ ３Ｇ１作为配基，淘筛噬菌体随机 １２肽库，经ＥＬＩＳＡ法鉴定后，对阳性克隆进行测序并与天然病毒蛋白Ｇ２及ＮＰ的序列进行同源性比较分析。结果 经３轮筛选，噬菌体显示有良好的富集效果。从第３轮筛选产物中，随机挑取２１个克隆进行ＥＬＩＳＡ结合实验。结果显示，９个克隆与 ｍＡｂ３Ｇ１有较强的特异结合活性；ＤＮＡ测序结果表明，具有保守性序列模式ＰＸ＿（１－２）ＨＸ＿（０－２）。该基序分别与Ｇ２～（９６）ＹＰＷＨＴＫＣＨＹ～（１０５）及 ＮＰ～（８１）ＰＹＧＶＥＰＧＤＨＬＫＥＲＳ～（９４）相似。结论 从噬菌体随机肽库中，筛选到能与ｍＡｂ ３Ｇ１特异性结合的一段短肽基序 ＰＸ＿（１－２）ＨＸ（０－２），其与 ＨＴＮＶ Ｇ２部分氨基酸的同源性较高，可能是与宿主细胞受体结合的表位，为进一步证实和研究ＨＴＮＶ的受体奠定了基础。
Aim To determine a epitope on HTNV that could bind to the re- ceptor on the host cells. Methods A type -specific mAb 3G1 could react to both HTNV glycoprotein G2 and nucleocapsid protein ( NP) and possessed fairly high neutralizing titer. Purified mAb 3G1 was used as the ligand for biopanning from a phage -displayed 12 amino acid peptide library, and then the binding capacity between phages and mAb 3G1 was examined by EHSA. The amino acid sequences of positive phage clones were compared with that of wild HTNV 76 -118 strain G2 and NP in homology respect. Results After 3 rounds of effective screening, 9 positive clones screened out of 21 clones were chosen and sequenced, we found a conservative motif PX(1-2) HX(0-2) H which was similar to G2^(96) PWHTAKCHY^(105) VFGVEPGDHLKERS^(94),re- spectively. Conclusion The clones reacting to mAb 3G1 can display the motif PX_( 1-2) HX_(0-2) H which shares the homologous amino acid sequence to G2. It indicates that the motif may be a epitope that can bind to the receptor on host cells . From this, a further research about receptor can be made.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2002年第1期73-75,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
军队医药卫生科研基金资助
NO.98D47
军队"十五"科研规划青年基金
NO.01 Q115
国家自然科学基金课题
NO.
