遗传 ›› 2003, Vol. 25 ›› Issue (4): 409-413.
龙华1;木下政人2 LONG Hua1;MASATO Kinoshita2
摘要: 从载体pBluescript SK+的Nde I位点处插入外源基因片段,构建了含有青鳉(Oryzias latipes)β-肌动蛋白(β-Actin)启动子和绿色荧光蛋白(GFP)标记基因的新表达载体。显微注射实验证实:在青鳉受精卵的单细胞期进行显微注射,标记基因的表达率和胚胎的存活率均较高。对嵌合体的转基因青鳉(F1)进行了两代的杂合体(F2和F3)筛选和一代的纯合体(F4)筛选,获得第4代转基因青鳉纯系品种。立体荧光显微镜检测结果表明:绿色荧光蛋白(GFP)基因是一种理想的标记基因。
Abstract:A new expression vector with the promoter of the medaka (Oryzias latipes) β-Actin promoter and green-fluorescent protein (GFP) gene was constructed based on Nde I site of pBluescript SK+.Microinjection experimentation proved that both expression rate of marker gene and the survival rate of transgenic embryos are high.The fourth generation of transgenic medaka,inbred variety,was obtained by heterozygote (F2 and F3) breeding of two generations and homozygote (F4) breeding of one generation based on mosaic (F1) of transgenic medaka.The detection results of stereo-fluorescence microscope show that green-fluorescent protein (GFP) gene is a kind of ideal marker gene.