摘要
目的分析不同刺激下外周血单个核细胞 (PBMC)中 L IGHT基因的表达 ,并克隆全长的人 L IGHT基因。方法按常规方法分离正常人的外周血单个核细胞后 ,培养于含 10 % FCS的 RPMI16 40中 ,并以不同浓度的 L PS、TNFα、IL-2、IFN- γ、PHA及 PMA刺激不同的时间 ,再以 RT- PCR法分析 PMBC内 L IGHT基因转录表达 ,并克隆相应的全长人L IGHT基因。结果 RT- PCR分析表明 :当用 L PS和 IFN- γ刺激 PMBC2 4h后 ,可诱导 L IGHT基因的明显表达 ,而单独应用IL - 2、TNFα、PHA和 PMA则不能诱导其表达。采用 PCR产物克隆方法 ,克隆了人 L IGHT基因 ,并经 DNA序列测定证实。结论本实验克隆得到了人 L IGHT基因 ,为进一步研究 L
ObjectiveTo analyze LIGHT gene expression in peripheral blood mononuclear cells (PBMCs) stimulated with different stimulators and to clone LIGHT gene Methods PBMCs were conventionally isolated and cultured in RPMI 1640,containing 10% FCS After stimulated with LPS,TNFα,IL 2,IFN γ,PHA and PMA ,the LIGHT gene expression in PBMCs was analyzed by RT PCR and LIGHT cDNA was cloned Results LIGHT expression in PBMCs could be detected by RT PCR in the group stimulated by IFN γ and LPS for 24 hours,however, it could not be detected in other groups stimulated with TNFα,IL 2,PHA and PMA respectively Then using PCR product cloning methods, LIGHT was cloned and sequenced Conclusion Human LIGHT gene was cloned successfully,laying the fundations for further research of LIGHT
出处
《免疫学杂志》
CAS
CSCD
北大核心
2001年第6期440-443,共4页
Immunological Journal
