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Construction of the plasmid pEGFP-N1-GGF2 and its expression in the rat brain
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF NEUROMEDICINE
- Issue:
- 10
- DOI:
- 10.3760/cma.j.issn.1671-8925.2007.10.001
- Key Word:
- 颅脑损伤;基因疗法;生长物质;脂质体;绿色荧光蛋白
Abstract： 目的 构建pEGFP-N1-GGF2质粒,观察其在大鼠脑组织中的表达.方法 采用RT-PCR方法从人胚胎脑组织总RNA中扩增出人GGF2全长序列,并克隆到增强型绿色荧光蛋白(EGFP)基因真核表达载体pEGFP-N1上,构建pEGFP-N1-GGF2质粒,通过脂质体将pEGFP-N1-GGF2质粒转染大鼠脑组织,荧光显微镜下观察GGF2融合蛋白在大鼠脑内表达的时间和空间分布特征.结果 成功构建了pEGFP-N1-GGF2表达载体,荧光显微镜观察可见,pEGFP-N1-GGF2表达载体转染6 h大鼠脑内即可见GGF2融合蛋白表达,3 d时融合蛋白表达最多.7 d时表达量稍有下降但仍高,14 d时表达量已明显下降.GGF2融合蛋白在脑内的表达以针道为中心向周围扩散,荧光信号可达皮层深部.结论 脂质体介导的pEGFP-N1-GGF2质粒能够在大鼠脑内表达GGF2融合蛋白.
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