欢迎您！东篱公司
退出
-
申报数据库
-
立项数据库
-
成果数据库
-
项目获奖数据库
中文摘要：
旨在构建内蒙古白绒山羊成纤维细胞生长因子Ⅴ（Fibroblast growth factor 5,FGF5）基因shRNA真核表达载体（pRNAT-shRNA）。以内蒙古白绒山羊胎儿成纤维细胞总RNA为模板,PCR扩增FGF5基因。设计并合成3条FGF5基因的靶向shRNA序列,分别插入pRNAT-U6.1/Neo载体中,构建重组干扰质粒载体pRNATshRNA1、pRNAT-shRNA2和pRNAT-shRNA3,用脂质体LipofectamineTM2000将各重组干扰质粒转染内蒙古白绒山羊胎儿成纤维细胞,Real time PCR检测FGF5mRNA表达量。结果表明:克隆获得813bp的目的基因,包含了完整的ORF,编码270个氨基酸。核苷酸序列及推导的氨基酸序列与绵羊FGF5基因（NM001246263.1）及氨基酸序列同源性均为99%。FGF5基因shRNA重组质粒经测序鉴定证明构建成功。重组表达质粒转染绒山羊胎儿成纤维细胞48h后可见绿色荧光表达;pRNAT-shRNA1组、pRNAT-shRNA2组和pRNAT-shRNA3组FGF5的mRNA表达水平均低于干扰空载体对照组（P【0.01）,pRNAT-shRNA2对FGF5的表达具有最佳的抑制效应。综上表明,成功构建pRNAT-U6.1/Neo-FGF5表达载体,为探讨FGF5在毛囊生长中的作用奠定了基础。
同期刊论文项目
同项目期刊论文
Molecular Characterization and Tissue-specific Expression of a Novel FKBP38 Gene in the Cashmere Goa
The novel mTOR inhibitor CCI-779 (temsirolimus) induces antiproliferative effects through inhibition
Molecular Cloning and Characterization of PI3K-p85α Gene from InnerMongolia Cashmere Goat (Capra hir
Direct Interaction between Ras Homolog Enriched in Brain and FK506 Binding Protein 38 in Cashmere Go
Overexpression of protein kinase B/AKT induces phosphorylation of p70S6K and 4E-BP1 ingoat fetal fib
Inhibitionof Mammalian Target of Rapamycin Complex 1 (mTORC1) Downregulates ELOVL1 GeneExpression an
Direct Interaction between Ras Homolog Enriched in Brain and FK506 Binding Protein38 inCashmere Goat
期刊信息
位置：