内蒙古白绒山羊VEGF164基因cDNA克隆及其组织表达特异性分析.pdf

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2011 2 VEGF164 cDNA 鲍文蕾 杨娇馥 李彬 刘俊娥 王潇 郭旭东 王志钢 侯鑫 刘东军 ( , 010021) : 旨在克隆内蒙古白绒山 血管内皮生长因子( vascu lar endothelial grow th factor, VEGF164)基因并分析其基本表达 模式采用 R -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 R - PCR方法进行组织表达检测获得了内蒙古白绒山 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 的 VEGF164( EU8576231)基因同源性为 99%, 相应的氨基酸序列同源性为 99%SMAR 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ProtCom p Version 90程序分析将其定位 于细胞外R -PCR检测表明, VEGF164基因在绒山 脑心脏睾丸胰腺脾肾和肺组织中均有表达 : 内蒙古白绒山 血管内皮生长因子 基因克隆 表达模式 Cloning and Characterization ofVEGF164 Gene cDNA and Analysis of Its Tissue-s ecific Ex ression in InnerM ongolia Cashmere Goat BaoW enlei Yang Jiaofu LiBin Liu June W ang Xiao Guo Xudong W ang Zhigang Hou X in L iu Dongjun (K ey Laboratory of MammalR eproductiveBiology and Biotechnology,M inistry of Education, College of Lfi eSciences, InnerMongolia niversity,H ohhot 010021) A bstrac:t he present study ami s at clon ing the CDS fragm ent ofVEGF164 gene cDNA in InnerM ongoliaCashm ere Goat and an- alyzing its tissue-specific expression. VEGF164 gene cDNA w as cloned by R -PCR. he nucleotide sequence w as analyzed by BLAS wh ile am ino acid sequencew as analyzed by online sofwt ares SMAR and Psite. he tissue-specific expression pattern ofVEGF164w as analyzed by sem -iquantitative R -PCR. he cloned VEGF164 gene cDNA w as 573 bp in length, includ ing a com plete ORF encod ing 190 am ino acids. he am ino acid sequence shares 99% identity w ith theOvis aries VEGF164 ( EU8576231). Analysisw ith SMAR suggested that the encoded protein