犬血管内皮细胞生长因子蛋白质和基因序列分析

Nucleotide Sequence and Expression of the Canine Vascular Endothelial Growth Factor

为构建和表达犬血管内皮细胞生长因子 (VEGF)奠定基础 ,从犬心脏组织提取总 RNA和蛋白质 ,利用逆转录 -聚合酶链反应方法 ,克隆 VEGF基因 ,将其重组入测序载体测序 ;采用 Western免疫印迹方法检测 VEGF蛋白质。结果发现 :犬心脏 VEGF c DNA存在 3种异构体 ,其序列长 6 35 bp、 5 6 3bp和 431bp,编码 188、 16 4和 12 0个氨基酸。犬 VEGF与人 VEGF相比 ,缺少第 8位氨基酸 -甘氨酸 ,同源性高达 94%。在犬心脏 Western免疫印迹方法检测到两种VEGF蛋白质的表达 ,分子量分别约为 46 ku和 30 ku。提示 :VEGF1 64 是犬心脏组织中主要的血管新生、血管张力和血管通透性调控因子。

The vascular endothelial growth factor (VEGF) gene was cloned by RT PCR from the canine heart for construction of recombinant VEGF. Total RNA and protein from the canine heart was extracted and degeneration primer was used to amplify by RT PCR. The VEGF protein in the tissue from heart was detected by using Western blot. Results showed that 3 cDNA, encoding the VEGF from the canine heart, were cloned and sequenced. Deduced amino acid sequence of these clones predicted 188 (VEGF 188 ), 164(VEGF 164 ) and 120 (VEGF 120 ) residues of VEGF, respectively. The canine VEGFs were one amino acid shorter as compared to human VEGF. Western blot analysis with poly clonal VEGF antibody showed about 46 ku and 30 ku VEGF protein species were expressed in the canine heart. It was suggested that VEGF 164 might be the predominant molecular species in the canine heart and probably play a critical role in angiogensis, vascular remodeling, and regulation of vascular tone and permeability.