红、黄、绿三种颜色荧光质粒导入大肠杆菌中的稳定性表达

Gene expression of red,yellow and green fluorescence plasmid stability after transferred in Escherichia coli

【目的】红、黄、绿3种颜色荧光蛋白质粒导入大肠杆菌的热稳定表达是实现质粒与受体菌杂交获得荧光标记菌的关键,荧光标记菌的构建,可极大地丰富荧光标记菌的颜色类别,为微生物示踪研究提供丰富的工具.【方法】采用大肠杆菌DH5α热激转导法构建荧光标记菌.【结果】黄色和绿色蛋白质粒在原液浓度基础上,稀释10倍进行热激转导,红色蛋白质粒原液直接进行热激转导,可将携带红、黄、绿3种颜色荧光蛋白的质粒DNA成功导入大肠杆菌.【结论】经PCR重复检测,条带明显并符合红、黄、绿3种颜色荧光质粒基因序列大小,3种颜色荧光质粒热稳定表达水平较高.

【Objective】The heat-stable expression of red,yellow and green fluorescent proteins after transferred into Escherichia coli is the key to realize the hybridization of plasmid and receptor bacteria to obtain fluorescent labeled bacteria.The construction of fluorescent labeled bacteria can greatly enrich the color of fluorescent marker bacteria,and provide abundant tools for microbial tracer research.【Method】DH5αwas used to construct fluorescent labeled bacteria.【Result】Yellow and green plasmid proteins were subjected to heat shock transduction with 10 times dilution,while the red plasmid protein was directly subjected to heat shock transduction without dilution.【Conclusion】PCR detection showed that the strips were obvious and accorded with the sequence sizes of 3 fluorescent plasmids.The plasmid DNA of red,yellow and green were transferred into Escherichia coli successfully with higher heat stable expression level.