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目的:探讨心脏发育相关基因ZNF394的功能。方法:构建ZNF394的真核表达载体EGPF-N1- ZNF394和重组子pCMV-tag2A-ZNF394,运用脂质体转染技术共转染COS-7细胞。整合外源基因,利用含有荧光素酶的报告基因检测新基因ZNF394的表达。结果:发现ZNF394蛋白定位在细胞核,该蛋白能抑制PFR-Luc和AP-1-Luc的活性,表明蛋白ZNF394参与了细胞分裂周期中信号途径的调控。同时,通过在实验室人工合成多态免疫新西兰大白兔,获得多克隆抗体,对人体胚胎进行抗体染色,发现新基因ZNF394在心脏中有表达。结论:以上实验证实了新基因ZNF374是一个与心脏发育有关的调控子,同时,对分析该基因功能的进一步研究打下了良好的基础。
Objective: To investigate the function of cardiac development related gene ZNF394. Methods: Eukaryotic expression vector EGPF-N1-ZNF394 and recombinant pCMV-tag2A-ZNF394 of ZNF394 were constructed and transfected into COS-7 cells by lipofection. The foreign gene was integrated and the reporter gene containing luciferase was used to detect the expression of the new gene ZNF394. Results: ZNF394 protein was localized in the nucleus. The protein inhibited the activity of PFR-Luc and AP-1-Luc, indicating that ZNF394 is involved in the regulation of signal pathways during cell division. At the same time, polyclonal antibodies were obtained by immunizing New Zealand white rabbits with artificial synthetic polymorphism in the laboratory, antibody staining was performed on human embryos, and the new gene ZNF394 was expressed in the heart. Conclusion: The above experiment confirmed that the new gene ZNF374 is a regulator of cardiac development, and at the same time laid a good foundation for the further study on the function of this gene.