福州大学学报（自然科学版）2018，Vol.46Issue(3)：438-444,7.DOI:10.7631/issn.1000-2243.17292
人源程序性死亡配体PD-L2蛋白的原核表达与抗血清制备
Prokaryotic expression and antiserum preparation of human programmed ligand PD-L2 protein
摘要
Abstract
The recombinant proteins hPD-L220-123( the major antigenic epitope region of human PD-L2) and hPD-L220-208(the extracellular domain of human PD-L2) were expressed in inclusion bodies. The two recombinant proteins were purified by Ni-NTA agarose resin and refolded. Then, the rat anti-serums against the recombinant proteins of hPD-L220-123 and hPD-L220-208 were prepared with titers of 1 : 106 measured by indirect ELISA. Western-blot experiments demonstrated that the prepared antise-rums can specifically recognize hPD-L2 protein. Finally, lung cancer tissues with high level expres-sion of hPD-L2 were used to assay the location of the prepared antiserums by IHC experiments, the results indicating that the membrane localization of the antiserum prepared by antigen of hPD-L220-123 protein was blur and weak;the membrane localization of the antiserum prepared by antigen of hPD-L220-208 protein had a specific localization, with less background and non-specific staining.关键词
人源PD-L2蛋白/主要抗原表位区/原核表达/抗血清Key words
human PD-L2 protein/major antigenic epitope region/prokaryotic expression/antiserum分类
生物科学引用本文复制引用
杨娟娟,刘照,夏菁潞,罗岭,俞博彤,孟春..人源程序性死亡配体PD-L2蛋白的原核表达与抗血清制备[J].福州大学学报（自然科学版）,2018,46(3):438-444,7.基金项目
国家自然科学基金资助项目(31700656) （31700656）
福建省自然科学基金资助项目(2017J05050) （2017J05050）
福州大学本科生科研训练计划(SRTP)资助项目(23119,23125) （SRTP）
福州大学贵重仪器设备开放测试基金资助项目(2017T025) （2017T025）
