新基因ZNF394的功能研究

Studies on the function of new gene ZNF394

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 discuss the function of cardiac development related gene ZNF394.Methods:The ZNF394 gene eukaryotic expression vector,pEGPF-N1-ZNF394 as well as recombinant pCMV-tag2A-ZNF394 were constructed,Application of liposome technique to transfection of COS-7 cells.The exogenous DNA integra- tion,then expression of ZNF394 gene were successively were detected out by using leuciferase assay with report gene.Result:Finding in the paper that ZNF394 protein localized at intranucleis,and the protein have the ability to inhibit the PFR-Luc and AP-1-Luc activities,it suggested the effects of the ZNF394 on signal pathways of cell division cycle.At the same,we discovered that the ZNF394 was expressed in heart;it was confirmed in hu- man embryo-staining using prepared with artificial synthetic peptide immune rabbit polyclone antibody in our laboratory.Conclusion:These findings strong documented that recombine ZNF394 is a regulator of cardiac func- tion further,simultaneously,it lays a basis for deeply analysing the gene function.