布氏鲳鲹VEGFA基因克隆与组织表达分析

Cloning and Tissue Expression Analysis of VEGFA Gene in Trachinotus blochii

血管内皮生长因子A (vascular endothelial growth factor A, VEGFA)主要生理功能是增加小静脉血管与微血管的通透性,是血管生成过程最重要的正向调控因子。本研究采用RACE技术克隆布氏鲳鲹VEGFA基因和实时荧光定量PCR方法对该基因进行组织分布表达分析。结果显示,该VEGFA基因cDNA序列全长1 470 bp,5’UTR、3’UTR长度分别为617 bp和208 bp,ORF为645 bp,共编码214个氨基酸,其中包括一个信号肽和两个N-糖基化位点。理论分子量为25.23 kD,等电点为8.91。同源性分析结果表明,布氏鲳鲹VEGFA基因与同属鲈形目鱼类高体鰤的同源性最高(96.1%)。荧光定量PCR分析显示,VEGFA在布氏鲳鲹组织中均有表达,其中肝脏表达水平最高,其次是心脏,在小脑、胃、肠表达较低,在肾脏组织中的表达量最低,说明布氏鲳鲹VEGFA基因在生理代谢以及血管生成过程中可能发挥重要调节作用。本研究结果为深入研究布氏鲳鲹VEGFA基因的生理代谢调控功能提供理论依据。

The main physiological function of vascular endothelial growth factor is to increase the permeability of venular vessels and microvessels,which is the most important positive regulator of angiogenesis.In this study,RACE and q PCR methods were used to study the cloning and tissue expression patterns of VEGFA gene.The results showed that the full-length c DNA of VEGFA gene was 1470 bp in length,5’UTR and 3’UTR were 617 bp and 208 bp in length,and the ORF was 645 bp.A total of 214 amino acids were included,including one signal peptide and two N-glycosylation site.The theoretical molecular weight is 25.23 kD and the isoelectric point is8.91.The results of homology analysis showed that the VEGFA gene of Trachinotus blochii had the highest homology with the homologous scorpionfish(96.1%).Quantitative real-time PCR analysis showed that VEGFA was expressed in Trachinotus blochii tissues,with the highest expression level of liver,followed by heart,lower expression in cerebellum,stomach and intestine,and lowest expression in kidney tissue,indicating Brinell.The VEGFA gene may have important regulatory functions during physiological metabolism and angiogenesis.The results of this study provide a theoretical reference for further study of the physiological metabolic regulation function of the VEGFA gene of Trachinotus blochii.