Molecular cloning, characterisation and functional analysis of the duck Forkhead box O3 (FOXO3) gene

作者：Gan, W.; He, H.; Li, L.^*

来源：British Poultry Science, 2016, 57(2): 143-150.

DOI：10.1080/00071668.2015.1135503

• SKELETAL-MUSCLE
• DRUG DISCOVERY
• IN-VITRO
• DIFFERENTIATION
• EXPRESSION
• CELLS
• BIOINFORMATICS
• ZEBRAFISH
• SURVIVAL
• EMBRYO

1. The Forkhead box O3 (FOXO3) transcription factor is a crucial regulator of cell fate that controls proliferation, apoptosis and differentiation. However, the role of FOXO3 regulation in duck myoblasts is not fully understood. @@@ 2. The aim of this study was to clone and determine the complete coding sequence (CDS) of the duck FOXO3 gene and to assess its function in myoblasts. @@@ 3. Primers specific for the predicted duck FOXO3 gene were designed using the public mallard duck reference sequence in GenBank. The CDS was cloned by RT-PCR and double digested to generate the expression vector pEGFP-N1-FOXO3. @@@ 4. Sequence analysis showed that the full-length FOXO3 CDS is 1467 bp, encoding 488 amino acids and is highly conserved across many bird species. Amino acid sequence analysis revealed a DNA-binding domain (aa 1-77). @@@ 5. Myoblast transfection with pEGFP-N1-FOXO3 showed that FOXO3 mRNA expression at 24 h was elevated in pEGFP-N1-FOXO3-transfected myoblasts compared to pEGFP-N1-transfected cells or controls. MRF4, MyoD, MyoG, Myf5 and PAX7 mRNA expression in the pEGFP-N1-FOXO3 group was lowest. However, myostatin (MSTN) and PAX3 mRNA expression did not differ. @@@ 6. These results suggest that FOXO3 plays a critical role in the proliferation and differentiation of duck myoblasts.

• 出版日期2016
• 单位四川农业大学