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目的 探讨细胞角蛋白 13(cytokeratin13,CK13)基因表达调控的机理 ,研究 CK13基因 5′旁侧不同基序对其转录活性的影响。 方法 采用分子克隆结合报告基因分析的方法 ,构建 CK 13基因 5′旁侧 5 13bp内不同基序与氯霉素乙酰转移酶 (chloramphenicol acetyltransferase,CAT)报告基因增强子载体p CAT的重组体 ,通过脂质体介导的转染技术导入 He L a细胞 ,检测各报告基因载体 CAT的相对活性。 结果 CK13基因 5′旁侧起始密码子 ATG上游 - nt.32 5～ - nt.2 0 7间 119bp中具有某种抑制子元件 ,-nt.2 0 6～ - nt.94间 113bp中具有某种增强子元件。 结论 CK13基因 5′旁侧 5 13bp内存在促进及抑制CK13基因表达的反应元件 ,进一步定位这些顺式反应元件并研究与之相互作用的反式作用因子 ,可望阐明 CK13基因表达调控及组织特异性表达的详细机理。
Objective To investigate the mechanism of cytokeratin 13 (CK13) gene expression and to investigate the effect of different motifs on the 5 ’side of CK13 gene in transcriptional activity. Methods The molecular cloning and reporter gene analysis was used to construct a 5 13 bp 5 ’flanking sequence of CK 13 gene and a recombined plasmid pCAT of chloramphenicol acetyltransferase (CAT) Liposome-mediated transfection technology was introduced into HeLa cells, and the relative activity of each reporter gene carrier CAT was detected. Results There were some suppressor elements in the 119bp upstream of the 5 ’- flanking start codon of CK13 gene - nt.32 5 ~ - nt.2 0 7, and 113bp in nt.2 0 6 ~ - nt 94 Some kind of enhancer element. Conclusion The 5 13 bp flanking 5 ’flanking 5’ of CK13 gene can promote and inhibit the expression of CK13 gene. To further locate these cis - acting elements and study their interaction with trans - acting factors, it is expected to clarify the regulation of CK13 gene expression and tissue specificity The detailed mechanism of sexual expression.