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CONSTRUCTION AND SCREENING EFFICIENT VEGF siRNA EXPRESSION VECTOR IN ZEBRAFISH
Authors
杨少丽
林秀坤
+3 more
王征
郑兰红
阎松
Publication date
31 December 2007
Publisher
Abstract
为探索小干扰RNA(small interfering RNA, siRNA)表达质粒在研究斑马鱼血管内皮生长因子(vascular endothelial growth factor,VEGF)基因调控网络中的应用,构建了4个以斑马鱼VEGF基因为靶点的siRNA表达载体pS1-VEGF、pS2-VEGF、pS3-VEGF及pS4-VEGF.通过显微注射的方法将载体导入1-2细胞期斑马鱼体内,于胚胎发育的48 h采用RT-PCR的方法检测VEGF基因的表达量,研究不同干扰序列对VEGF基因表达的干涉作用.结果显示,成功地构建了siRNA表达载体.针对不同位点的寡核苷酸序列抑制VEGF基因表达的效率有显著差异,其中注射了pS1-VEGF的胚胎出现了心包膜水肿、血流速度减慢、循环红细胞堆积等症状,同时肠下静脉、节间血管以及其它血管出现不同程度的发育缺陷.实验结果说明,pS1-VEGF可引起斑马鱼胚胎血管发育缺陷
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Last time updated on 18/09/2018