绿色荧光蛋白基因在转基因金鱼中的整合与表达

Integation and Expression of Green Fluorescent Protein Gene Goldfish

通过分子克隆技术将鲤鱼β—肌动蛋白启动子(carpβ-actinpromoterA)与编码绿色荧光蛋白(GFP)的cD NA融合构建成能在真核生物体内表达的质粒载体pAGFP。质粒pAGFP经PvuⅠ线性化后采用显微注射法导入金鱼受精卵,在胚胎发育初期,经紫外光激发能观察到少数胚胎发绿色荧光。在通过PCR实验检测出的5个阳性个体染色体DNA中,经点杂交实验证实有2个个体染色体基因组上整合了GFP基因。

GFP cDNA gene was subcloned to carp β-action promotor,then cxpression vector pAGFP was constructed,Vector pAGFP was injected into embryo of Goldfish in 1-2 stage after digestion with Pvu Ι.During the development of embryo,we can observe a few embryo emit Green Fluorescent Protein under UV light.In experiment,5 individuals can be applied a specific molecular band via per,and 2 individuals were comfirmed by Dot blot which have been integrated in Goldfish genome.