摘要
目的分析不同刺激下外周血单个核细胞(PBMC)中THANK基因的表达，并克隆全长的人THANK基因。方法采用将常规方法分离的外周血单个核细胞(PBMC)培养于含100ml/L FCS的RPMI 1640中，并以不同浓度的LPS、TNF-α、IL-2、IFN-γ、PHA及PMA刺激不同时间。以RT-PCR法，分析PMBC中THANK基因的转录表达，并克隆相应的全长人THANK基因。结果RT-PCR分析表明，当用IFN-γ刺激PMBC 72h后，可诱导THANK基因明显表达;而IL-2、LPS、TNF-α、PHA和PMA则不能诱导其表达。采用PCR产物克隆的方法，克隆了人THANK基因，并经DNA序列测定证实。结论克隆得到了人THANK基因，为进一步研究THANK的功能打下了基础。
Aim To analyze THANK gene expression in peripheral blood mononuclear cells(PBMC) stimulated with different stimulators and to clone whole-length human THANK gene. Methods PBMC were conventionally isolated and cultured in RPMI1640 containing 10％FCS. After stimulated with LPS,TNF-α,IL-2,IFN-γ,PHA or PMA,the THANK gene expression in PBMCs was analyzed by RT-PCR and THANK cDNA was cloned. Result RT-PCR detection showed that THNAK gene expressed in PBMCs after stimulated with interferon-γfor 3 days, whereas THANK'expression could not be detected after stimulated with LPS,TNF-α,IL-2,IFN-γ,PHA or PMA respectively. Then THANK gene was cloned by cloning PCR product and sequenced. Conclusion Human THANK gene is cloned successfully, thus providing the possibility for further research of THANK'function.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第4期304-306,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助
No. 39770694
