内蒙古白绒山羊VEGF164基因cDNA克隆及组织表达特异性分析

Cloning and Characterization of VEGF164 Gene cDNA and Analysis of Its Tissue-specific Expression in Inner Mongolia Cashmere Goat

旨在克隆内蒙古白绒山羊血管内皮生长因子(vascular endothelial growth factor,VEGF164)基因并分析其基本表达模式。采用RT-PCR技术克隆基因,将得到的基因cDNA序列及其编码的氨基酸序列进行生物信息学分析。利用半定量RT-PCR方法进行组织表达检测。获得了内蒙古白绒山羊VEGF164基因编码区cDNA全长序列,扩增片段全长573 bp,包含了完整的ORF,编码190个氨基酸残基。核苷酸序列与绵羊的VEGF164(EU857623.1)基因同源性为99%,相应的氨基酸序列同源性为99%。SMART程序分析表明,ORF编码的蛋白质具有信号肽序列及血小板衍生和血管内皮生长因子家族(PDGF,VEGF)结构域。Psite程序分析表明,有1个蛋白激酶C磷酸化位点,4个酪蛋白激酶磷酸化位点。ProtComp Version 9.0程序分析将其定位于细胞外。RT-PCR检测表明,VEGF164基因在绒山羊脑、心脏、睾丸、胰腺、脾、肾和肺组织中均有表达。

The present study aims at cloning the CDS fragment of VEGF164 gene cDNA in Inner Mongolia Cashmere Goat and analyzing its tissue-specific expression.VEGF164 gene cDNA was cloned by RT-PCR.The nucleotide sequence was analyzed by BLAST while amino acid sequence was analyzed by online softwares SMART and Psite.The tissue-specific expression pattern of VEGF164 was analyzed by semi-quantitative RT-PCR.The cloned VEGF164 gene cDNA was 573 bp in length,including a complete ORF encoding 190 amino acids.The amino acid sequence shares 99% identity with the Ovis aries VEGF164（EU857623.1）.Analysis with SMART suggested that the encoded protein contained a signal peptide and a Platelet-derived and vascular endothelial growth factors（PDGF,VEGF） family domain.Analysis with Psite indicated a Protein kinase C phosphorylation site and four Casein kinase II phosphorylation sites in this protein.Analysis with ProtComp Version 9.0 suggested this protein to be extracellular（secreted）.VEGF164 gene was shown to express in brain,heart,testis,pancreas,spleen,kidney and lung.