黄颡鱼bmp2a与bmp4基因启动子的克隆及分析

Molecular cloning and analysis of bmp2a and bmp4 promoters in yellow catfish Pelteobagrus fulvidraco

为研究黄颡鱼bmp2a与bmp4基因的功能及其相关转录因子的调控网络,采用RLM-5′RACE方法确定bmp2a与bmp4基因的转录起始位点,通过hiTAIL-PCR方法克隆bmp2a与bmp4基因启动子序列并运用生物信息学方法预测启动子序列上的关键转录因子结合位点。结果显示,bmp2a与bmp4基因的转录起始位点分别位于bmp2a与bmp4基因编码区上游的391bp与351bp处。克隆bmp2a与bmp4的1 830bp、1 962bp启动子序列,在启动子序列的基础上预测出AP1、SP1、E-box、GATA1、CREB、PPARγ、SOX5、SOX6和SOX9等转录因子的结合位点,提示这些转录因子对bmp2a与bmp4基因的转录调控发挥潜在的重要作用。

To explore the function of the bmp2a and bmp4 genes and regulatory networks of the related transcription factors in yellow catfish Pelteobagrus fulvidraco,RLM-5′RACE method was used to identify the transcription start site(TSS)of bmp2a and bmp4.Then,the promoters of bmp2a and bmp4 were cloned by hiTAIL-PCR method and a cluster of putative binding sites of several transcription factors were identified by bioinformatics analysis.The results showed that the TSS of bmp2a and bmp4 were located at 391 bp and 351 bp upstream of the coding sequence,respectively.Then 1 830 bp and 1 962 bp upstream of the TSS of bmp2a and bmp4 were cloned and key binding sites of several transcription factors,such as AP1,SP1,GATA1,CREB,PPARγ,SOX5,SOX6 and SOX9,were predicted,suggesting that these transcription factors may play crucial roles in the transcriptional regulation of the bmp2a and bmp4 genes.