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JNKs是丝裂原活化蛋白激酶(MAPK)家族重要成员,参与应激反应和动物体轴的构建。为了进一步了解JNK家族在动物发育中的功能,首次分离和克隆了金鱼jnk3基因,研究了jnk3在金鱼和斑马鱼组织特异性和胚胎发育特异性表达状况。金鱼jnk3基因cDNA总长1 794 bp,其中阅读框1 293 bp,编码430个氨基酸。对jnk3基因序列及其推导的氨基酸序列进行同源性分析,结果显示,jnk3基因在脊椎动物较为保守,金鱼jnk3基因核苷酸序列与斑马鱼、人的同源性分别为94.1%和80.7%,金鱼jnk3基因氨基酸序列与斑马鱼、人的同源性分别为99.7%和93.4%。jnk3基因在鱼类成体中的表达存在着显著的组织差异,在金鱼和斑马鱼脑和精巢组织的表达具专一性,此外在斑马鱼心脏中也检测到jnk3的表达。RT-PCR检测结果显示,在鱼类胚胎发育中,神经胚期开始检测到jnk3基因mRNA表达,从神经胚到心跳期胚胎,jnk3基因表达水平逐渐增高,此后直至出膜一直保持较高的表达水平。研究表明,jnk3基因可能在鱼类后期的胚胎发育和成体脑、精巢与心脏发育中具有重要作用。
JNKs are important members of the mitogen-activated protein kinase (MAPK) family and are involved in stress response and body axis building. In order to further understand the function of JNK family in animal development, the jnk3 gene was isolated and cloned for the first time to study tissue-specific and embryo-specific expression of jnk3 in goldfish and zebrafish. The cDNA of jnk3 gene was 1 794 bp in length, including 1 293 bp in reading frame and 430 amino acids. The homology analysis of jnk3 gene and its deduced amino acid sequence showed that jnk3 gene was more conserved in vertebrates, and the homologies of the jnk3 gene of goldfish with zebrafish and human were 94.1% and 80.7% , Goldfish jnk3 amino acid sequence and zebrafish, human homology were 99.7% and 93.4%. There was significant difference in the expression of jnk3 gene between adult fish and zebrafish brain. The expression of jnk3 was also detected in zebrafish brain and testes. The results of RT-PCR showed that in the process of fish embryo development, the expression of jnk3 gene was detected at neurogenetic stage. The expression of jnk3 gene gradually increased from embryo to heartbeat embryo, and maintained high expression Level. Studies have shown that the jnk3 gene may play an important role in late embryonic development and adult brain, testis and heart development in fish.