申请人克隆了一个心脏高表达的基因Asb5，但目前尚无该基因在动物心脏发育中功能的有关研究报道，申请人前期研究结果提示Asb5基因可能与心脏的发育有关。本研究拟首先构建Asb5转基因和基因敲除斑马鱼模型，然后利用斑马鱼动物模型和细胞系对Asb5基因在心脏器官发育中的作用及其分子调控机制进行研究。主要采用胚胎原位杂交、免疫组化、Western blot、Pull down、染色质免疫沉淀等方法，研究Asb5基因与心脏器官发育的关系、其在心脏发育过程中的作用、可能参与的信号途径以及转录调控的机理。本研究对阐明Asb5基因在动物心脏器官发育中的作用及调控途径具有十分重要的理论意义。

The applicant has cloned a zebrafish heart high-expression gene Asb5. However, the function of this gene in the heart development has not been reported yet. The results of applicant preliminary study have showed that the Asb5 gene might be involved in heart development. In this project, Asb5 gene knockout and transgenic experimental models will be established firstly. The role and molecular mechanism of Asb5 in heart development will be studied then using those zebrafish experiment model and cell line as well. The following methods such as in situ hybridization, immunohistochemistry, Western blot, Pull down and Co-IP etc will be used to study the relation between Asb5 gene and the occurrence of heart development, its role in the course of heart development, the possible signaling pathway and the potential transcriptional regulation mechanism of heart development. This research might clarify the role of Asb5 gene and its mechanism in animal heart development.