绿色荧光蛋白的基因克隆和表达研究 （湖北师范学院生命科学学院 生物科学0802 班 湖北 黄石 435002） 摘 要：目的：研究绿色荧光蛋白（Greed Flu orescent Protein，GFP）基因的基因克隆及在大肠杆菌中的表达。方法：通过分别将DH-5α (pEGFP-N3)和DH-5α(pET-28a)提取质粒、酶切并连接形成重组质粒pET-28a-GFP，将重组质粒导入 E.co li DH-5α 感受态细胞中进行转化,通过限制性核酸内切酶Not I 与 Bam H1和PCR 对所建质粒进行分析鉴定后, 通过转化的方法把含绿色荧光蛋白（GFP）外源基因转入大肠杆菌体 BL-21内进行表达，再用 IPTG诱导 GFP 基因表达，可以看到显现绿色，判断 GFP 基因在大肠杆菌中成功表达。 结果：结果显示构建的重组质粒pET-28a-GFP 在E.co li 中成功表达。 关键词：绿色荧光蛋白；质粒重组；原核表达；诱导表达 中图分类号：Q53 Studies On Cloning and Expression of Green Fluorescent Protein gene Abstract: Objective:Studies indicated that the cloning and expression of the GFP gene in the E.co li. Methods: Extract the plasmid of the DH-5α(pEGFP-N3) and DH-5α (pET-28a). Then cutting by enzyme and connecting the two plasmids to form pET-28a-GFP recombined plasmid. The recombinant plasmid confirmed by restriction enzyme and PCR transfected into E.co li DH-5α to ensure the expression of green fluorescent protein. Guiding the recombined plasmid, which contains exogenous genes of GFP into E.co li for expression, through transformative method. The expression of GFP gene can be induced by the IPTG and then we can see green. Results: The results suggest that pET-28a-GFP recombined plasmid has successfully expressed in E.co li. Keywords: Ged Fluorescent Protein; Recombined Plasmid; Prokaryote Expression; Induced Expression 湖北师范学院分子生物学实验论文 1 绿色荧光蛋白的基因克隆和表达的研究 引言 随着分子生物学和基因工程技术的迅速发展和广泛应用, 人们根据自己的意愿有目的、有计划、有根据、有预见地将外源基因导入动物细胞内, 使外源基因进行表达、阐明基...
