GATA4/5/6基因克隆及其在出生后鸭心脏和肝脏中的发育性表达模式

Cloning of GATA4/5/6 Genes and Their Expression Patterns in Duck Hearts and Livers during Postnatal Development

旨在克隆和分析鸭内脏器官发育调控关键基因GATA4/5/6编码区,并初步揭示其mRNA表达量与出生后鸭心脏和肝脏发育的关系。本研究对鸭GATA4/5/6基因进行PCR扩增和克隆,对得到的编码区进行序列分析,采用qRT-PCR检测它们在0、2、4、6、8周龄鸭心脏和肝脏中的表达量,并对表达量与心脏和肝脏重量及器官指数进行相关分析。结果,获得鸭GATA4/5/6基因完整编码区序列,长度依次为1 242、1 182、1 173 bp。鸭GATA4/5/6均存在位于N端和C端的2个锌指结构域以及1个GATA-N转录激活结构域,3个基因间GATA-N转录激活域的氨基酸一致率低,而2个锌指结构在GATA4/5/6中共有区域的保守性很高,但GATA5的C端锌指结构和GATA6的N端锌指结构分别缺失了11和15个氨基酸。qRT-PCR结果显示,GATA4/5/6均在0~8周龄鸭心脏和肝脏中一直维持较高表达水平,但不同基因间的发育性表达模式明显不同,提示可能存在功能差异。进一步的相关性分析表明,鸭心脏和肝脏的重量与GATA4表达量极显著相关(P<0.01)。综上,本试验初步揭示GATA4/5/6在出生后鸭心脏和肝脏中持续性表达并具有各自特异的表达模式,并且可能由于GATA4具有更完整的锌指结构域使得其与心脏和肝脏发育关系尤为密切。

The aim of this study was to clone and analyze the complete coding region sequences of GATA4/5/6, the key genes regulating the development of duck internal organs. The relationship between mRNA expression levels of GATA4/5/6 at 0, 2, 4, 6,8 weeks old and the development of heart and liver of duck was preliminarily revealed in duck. PCR amplification and cloning were performed on the GATA4/5/6 genes of ducks, and sequence analysis was performed on the coding regions. qRT-PCR was used to detect GATA4/5/6 expression in heart and liver of ducks at 0, 2, 4, 6, 8 weeks old. The correlationship analysis between expression level of GATA4/5/6 and heart and liver weight and organ index was performed. The complete coding region sequences of duck GATA4/5/6 genes were obtained by amplification, with the length of 1 242, 1 182 and 1 173 bp, respectively. Further analysis showed that GATA4/5/6 of ducks had 2 zinc finger domains located at the N-terminal and C-terminal and one GATA-N domain. The amino acid consistency rate of GATA-N transcriptional activation domain was low among the 3 genes. The common areas of the 2 zinc finger structures in GATA4/5/6 were higher, but the zinc finger structure at the C-terminal of GATA5 and the zinc finger structure at the N-terminal of GATA6 were the sequence deleting 11 and 15 amino acids, respectively. qRT-PCR results showed that GATA4/5/6 maintained a high expression level in the heart and liver of 0-8 weeks old ducks. But, the developmental expression patterns between different genes and tissues were significantly different. This phenomenon indicated that GATA4/5/6 might have different functions. Further correlation analysis showed that the weight of duck heart and liver was significantly correlated with GATA4 expression level(P<0.01). In conclusion, the present study preliminarily revealed that the sustained and specific expression of GATA4/5/6 might be related to the development of heart and liver in postnatal ducks. Moreover, GATA4 might be particularly closely related to the development of heart and liver due to its more complete zinc finger domain.