Molecular cloning, expression analysis and miRNA prediction of vascular endothelial growth factor A (VEGFAa and VEGFAb) in pond loach Misgurnus anguillicaudatus, an air-breathing fish

作者：Luo, Weiwei; Liang, Xiao; Huang, Songqian; Cao, Xiaojuan^*

来源：Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology, 2016, 202: 39-47.

DOI：10.1016/j.cbpb.2016.07.006

• Misgurnus anguillicaudatus
• Vascular endothelial growth factor A
• Molecular cloning and expression
• MiRNAs prediction

Vascular endothelial growth factor A (VEGFA) is the most studied and the best characterized member of the VEGF family and is a key regulator of angiogenesis via its ability to affect the proliferation, migration, and differentiation of endothelial cells. In this study, the full-length cDNAs encoding VEGFAa and VEGFAb from pond loach, Misgurnus anguillicaudatus, were isolated. The VEGFAa is constituted by an open reading frame (ORF) of 570 bp encoding for a peptide of 189 amino acid residues, a 639 bp 5'-untranslated region (UTR) and a 2383 bp 3' UTR. The VEGFAb is constituted by an ORF of 687 bp encoding for a peptide of 228 amino acid residues, a 560 by 5' UTR and a 1268 bp 3' UTR. Phylogenetic analysis indicated that the VEGFAa and VEGFAb of pond loach were conserved in vertebrates. Expression levels of VEGFAa and VEGFAb were detected by RT-qPCR at different development stages of pond loach and in different tissues of 6-month-old, 12-month-old and 24-month-old pond loach. Moreover, eight predicted miRNAs (miR-200, miR-29, miR-218, miR-338, miR-103, miR-15, miR-17 and miR-223) targeting VEGFAa and VEGFAb were validated by an intestinal air-breathing inhibition experiment. This study will be of value for further studies into the function of VEGFA and its corresponding miRNAs, which will shed a light on the vascularization and accessory air-breathing process in pond loach.

• 出版日期2016-12
• 单位华中农业大学