【摘要】 通过体外重组,将绿色荧光蛋白基因编码序列克隆到鲤鱼肌动蛋白基因启动子下游,构建成能在鱼体内表达绿色荧光蛋白的重组分子。用限制性内切酶PvuI酶切线性化后,通过显微注射法导入金鱼受精卵内,在转化后48h,可观察到绿色荧光,经PCR初步筛选后,对显阳性个体进行Southern杂交检测,结果表明,转化个体表现出较强的杂交信号。这说明绿色荧光蛋白基因能在金鱼体内整合、表达。更多还原

【Abstract】 The coding sequence of green fluorescent protein gene was cloned under the control of carp β-actin gene promoter. A fish expression of green fluorescent protein gene recombinant was constructed. The recombinant gene was transferred into fertilized zygotes of goldfish (Carassius auratus) via microinjection after PvuⅠlinearing. The green fluorescence was visualized within 48 h following transformation. The transformation and expression of green fluorescent protein was demonstrated by PCR and Southern blotting. In Southern hybridization analysis, transformations got positive signals. The results indicated that the green fluorescent protein gene had integrated into goldfish genome.更多还原