犬血管内皮细胞生长因子蛋白质和基因序列分析

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 cDNA存在3种异构体,其序列长635 bp、563 bp和431bp,编码188、164和120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VEGF164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.

Nucleotide Sequence and Expression of the Canine Vascular Endothelial Growth Factor

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.