抗禽白血病病毒p27蛋白线性抗原表位的鉴定  

Identification of a linear B-cell epitope specific for avian leukosis virus p27 protein

作　　者：李晓菲[1] 朱海波[2] 高奇[1] 王琦[1] 孙佳善[2] 高玉龙[1] 祁小乐[1] 王永强[1] 高宏雷[1] 王笑梅[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽免疫抑制病科技创新团队,黑龙江哈尔滨150001 [2]哈尔滨动物用生物制品国家工程研究中心有限公司,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2015年第8期632-635,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家现代农业产业技术体系(nycytx-42-G3-01);国家自然科学基金(31372437;31201923)

摘　　要：为鉴定禽白血病病毒(ALV)p27蛋白中的线性抗原表位,本研究以原核表达的重组p27蛋白免疫BALB/c小鼠,利用淋巴细胞杂交瘤技术,将免疫鼠脾细胞与SP2/0细胞融合,利用间接ELISA方法筛选纯化获得一株能够稳定分泌特异性单克隆抗体(MAb)的杂交瘤细胞株。Western blot试验结果显示,该MAb可以识别ALV的群特异性抗原p27蛋白。间接免疫荧光试验结果显示,MAb与A、B和J亚群ALV均呈阳性反应。对p27蛋白进行截短表达,利用western blot和间接ELISA进行表位鉴定,确定了MAb所识别的抗原表位位于129LVAITASALQAFRE142。氨基酸序列比对结果显示,本研究所鉴定出的p27表位在A、B和J亚群ALV之间高度保守。本研究为ALV检测方法的建立及其免疫学功能的研究提供了有利的工具。To identify the linear epitope for avian leukosis virus （ALV） p27 protein, the spleen cells from the BALB/c mouse immunized with the purified recombinant p27 proteins were fused with the SP2/0 cells. Then one hybridoma cell stably secreting monoclonal antibody （MAb） against the p27 protein was seaned by indirect ELISA. The result showed that the MAb was able to recognize the ALV p27 protein of ALV-A, ALV-B and ALV-J. In addition, a series of partially overlapping peptides for p27 encoding sequences were expressed and screened by MAb. Results showed that the linear epitope on the p27 protein was located in the 129LVAITASALQAFRE142. Alignment analysis of amino acid sequences indicated that the identified epitope was highly conserved among ALV-A, ALV-B and ALV-J. The prepared MAb and the identified epitope might provide valuable tool for the development of ALV immunodiagnostic approaches and the study of the function of the p27 protein.

关 键 词：禽白血病病毒 P27蛋白 单克隆抗体 抗原表位 

分 类 号：S852.65[农业科学—基础兽医学]