CKLF基因与CKLFSF1基因间的顺式调控元件  

A novel cis-acting enhancer element between CKLF and CKLFSF1 genes

作　　者：徐明旭[1] 赖茂毅[1] 夏东岚[1] 王莺[1] 张颖妹[1] 王露[1] 韩文玲[1] 

机构地区：[1]北京大学人类疾病基因中心,北京100083

出　　处：《细胞与分子免疫学杂志》2003年第3期276-278,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目 (No .30 1 70 871 )

摘　　要：目的 :探讨CKLF基因与CKLFSF1基因间序列 (CCS)的调控作用。方法 :运用PCR技术扩增CCS ,并将此片段插入含有荧光素酶 (luciferase)报告基因载体 pGL3 basic ,以及 pGL3 SV4 0启动子中 ,构建受其调控的荧光素酶报告基因载体。应用脂质体介导基因转染技术 ,将 4种重组质粒转染Hela细胞 ,进行瞬时表达分析。结果 :在pGL3 basic和 pGL3 basic CCS质粒中的报告基因luciferase均无表达 ,但将CCS片段插入至 promoter上游后 ,荧光素酶活性增加近 1倍。 结论 :CKLF与CKLFSF1基因间的序列不具有启动子活性 。AIM: To explore the roles of the CKLF gene and CKLFSF1 gene sequence (CCS) in transcriptional regulation. METHODS: The target gene fragment was amplified by PCR and then inserted into pGL3 basic and pGL3 SV40 containing luciferase reporter vector gene to construct pGL3 basic CCS and pGL3 SV40 CCS. Using liposome mediated method, four recombinant plasmids were respectively transfected into Hela cells. Transient expression was analyzed. RESULTS: The luciferase assay indicated that the no luciferase activity was detected in Hela cells transtected with pGL3 basic and pGL3 basic CCS. However, the luciferase activity was doubled when pGL3 SV40 CCS was transfected into Hela cells. CONCLUTION: The CCS has no promoter activity, whereas some important cis acting enhancer elements which modulate its downstream gene expression may exist within this sequence.

关 键 词：转录因子 调控 荧光素酶 

分 类 号：Q78[生物学—分子生物学] R392-33[医药卫生—免疫学]