摘要
目的构建GnRH/TRS与绿色荧光蛋白(GFP)的融合基因并表达。方法利用DNA重组技术,将GnRH/TRS片段克隆至真核表达载体pEGFP-C1转化DH5α,重组体质粒pEGFP-C1-GnRH/TRS用LipoGen脂质体进行转染至Hela细胞,核酸序列测定和Western印迹分析基因表达,激光共聚焦荧光显微镜观察活细胞内荧光布局。结果重组子pEGFP-C1-GnRH/TR成功构建。激光共聚焦荧光显微镜观察结果表明,GnRH/TRS-GFP融合基因的瞬间和稳定表达均获得了相同结果。结论GnRH/TRS-GFP融合蛋白具有GFP的自发荧光特性,且不影响GnRH/TRS分子在细胞内的正确表达。
Objective To constructe A fusion gene composed of green fluorescent protein （GFP）and GnRH/TRS DNA then express .Methods Clone GnRH/TRS into eucaryotic expression vector pEGFP-C1 by recombinant DNA technique.then transform to DH5 α then pEGFP-C1-GmR.H/TR transfect into Hela by LipoGen.Snucleotid sequence and Western blot analysis were used to evaluate the expression of the fused gene. Laser-scanning confocal microscopy was used to monitor the distribution of green fluorescence. Results Recombinant plasmid pEGFP-C1-GnRH/TR was successfully constructed Laser-scanning confocal microscopy clearly revealed that GnRH/TRS-GFP can be transiently and stably expressed in mammalian cells. Conclusion The GFP tag does not interfere with the natural assembly and transport of GnRH/TRS molecule.
出处
《生物技术通报》
CAS
CSCD
2008年第1期122-123,155,共3页
Biotechnology Bulletin
关键词
GNRH
转运肽
绿色荧光蛋白
Gonadotropin releasing hormone
Transporter
Green fluorescent protein
