摘要
PD-1分子是一种重要的免疫调控因子,目前对其自身表达调控尚未有系统的研究.对PD-1启动子区域进行分析,克隆构建了含PD-1基因上游约2kb范围内含4种不同长度调控序列的双荧光素酶表达载体.通过FACS检测发现,小鼠T淋巴瘤EL4细胞稳定表达PD-1,而骨髓瘤Sp2/0-Ag细胞则在佛波酯(phorbol 12-myristate 13-acetate,PMA)和离子霉素(ionomycin,IO)诱导后才表达PD-1.4种双荧光素酶报告系统在上述两种细胞中检测PD-1启动子各区段活性显示,-227～+49bp区域含有PD-1核心启动子,上游-1127～-716bp含有较强正性调节元件,而在-1685～-1128bp、-715～-228bp两个区域含有负性调节元件,这种正负调控区交错的启动子活性现象说明了PD-1基因表达调控的复杂性,这些结果为PD-1基因的表达调控提供了结构基础和依据.
Extensive studies have been performed on function of PD-1 in immunological regulation, while as so far, studies on the exact regulation mechanism of PD-1 expression have not been reported. PD-1 expression in EL4 and Sp2/0-Ag cell lines was detected with PE-Anti Mouse-PD-1 antibody through FACS. Genomic DNA from C57BL/6J mouse was used to produce different lengths of PD-1 promoter fragments. The PCR products were cloned into the luciferase reporter vector pGL3-Basic and co-transfected with pRL-SV40 into EL4 ...
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2010年第5期527-533,共7页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金资助项目(30571650
30901315)
国家重点基础研究发展计划(973)资助项目(2006CB504304)
国家高技术研究发展计划(863)资助项目(2006AA02Z462)~~
关键词
PD-1
启动子鉴定
转录调控分析
PD-1
promoter identification
transcriptional regulation
