Fisher S A, Walsh K, Forehand C J
Department of Medicine, University of Vermont, Burlington, USA.
J Mol Cell Cardiol. 1996 Jan;28(1):113-22. doi: 10.1006/jmcc.1996.0011.
To study transcriptional regulation during early stages of cardiogenesis, stage 10-17 chicken embryo hearts were transfected efficiently within the intact embryo by application of plasmid DNA complexed to liposomes. Viral regulatory sequences and the skeletal alpha-actin, SERCA2 and ANF promoters activated expression of reporter genes in the primitive heart tube. Deletion and mutation analysis of the skeletal alpha-actin promoter revealed the importance of CArG cis-regulatory elements in enhancing transcription of the gene during early heart development. These results demonstrate the utility of this method for the identification of gene regulatory elements that specify the cardiac phenotype during early stages of heart morphogenesis.
为了研究心脏发生早期阶段的转录调控，通过应用与脂质体复合的质粒DNA，在完整胚胎内对第10 - 17阶段的鸡胚心脏进行了高效转染。病毒调控序列以及骨骼肌α - 肌动蛋白、肌浆网钙ATP酶2（SERCA2）和心钠素（ANF）启动子激活了原始心管中报告基因的表达。骨骼肌α - 肌动蛋白启动子的缺失和突变分析揭示了CArG顺式调控元件在心脏早期发育过程中增强该基因转录的重要性。这些结果证明了该方法在鉴定心脏形态发生早期阶段决定心脏表型的基因调控元件方面的实用性。
