摘要
探讨趋化素样因子超家族成员 1,2基因 (CKLFSF1基因与CKLFSF2基因 )间的短序列对其下游基因表达的调控作用 .运用PCR技术扩增CKLFSF1基因与CKLFSF2基因间的序列 ,将此片段插入含有萤光素酶 (luciferase)报告基因载体上 .以磷酸钙介导基因转染技术 ,将重组质粒以及阴性和阳性对照组质粒转染到HeLa细胞 ,进行瞬时表达分析 .在pGL3 Basic质粒中的报告基因萤光素酶无表达 ,但将CKLFSF1与CKLFSF2基因间的序列插入到启动子上游或下游后 ,显著抑制其下游基因的表达 ,萤光素酶活性明显降低 .结果提示 ,CKLFSF1与CKLFSF2基因间的序列不具有启动子活性 。
The roles of the chemokine\|like factor super family 1 and 2 ( CKLFSF 1 and CKLFSF 2) gene interval sequence was explored in transcriptional regulation. The target fragment was obtained by PCR amplification and inserted into pGL3 luciferase reporter vector. Using calcium phosphate precipitation mediated transient expression approach,the recombinant plasmids were transfected into HeLa cells respectively. The luciferase assay indicated that the luciferase activity was not detected when pGL3 Basic was transfected into HeLa cells. Nevertheless, the luciferase activity was markedly decreased when test plasmid was transfected into HeLa cells. The results suggested that the CKLFSF 1 and CKLFSF 2 gene interval sequence had no promoter activity, whereas some important cis acting silencer elements for the gene expression were localized within this sequence.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2003年第3期349-353,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金 (No.3 0 170 871)
国家 863高技术研究和发展计划 (No.2 0 0 1AA2 15 0 61)资助~~
