Abstract:

Objective To establish an effectively transgenic technique for chicken embryo tectum electroporation by a self-made electrode. Methods Each embryo was precisely injected 0.25-0.5μl pCAGGS-green fluorescent protein (GFP)plasmid (0.5g/L) in the opitc tectum, and then preformed in vivo electroporation in a specific time and position using a self-made electrode as test group and an original electrode as control (60 eggs for earch group), under the conditions of 18 volt, each pulse 60ms, interval 100ms, and 6 times conditions. After electroporation, we analyzed the survival and positive ratio of embryo at E6-E12 using stereo fluorescence microscope to observe the reporter gene GFP expression and to judge the efficiency of transfection. Results The survival ratio of chicken embryo whose tectum was electroporated by the self-made electrode reach to 89% after 24 hours and 35% at E12, which were 48.3% and 600% times higher than original electrode. Conclusion This study demonstrates that the self-made electrode can cause tiny damage and ensure the higher survival ratio of electroporated chicken embryos so that it provides an effective transformation technique for in vivo electroporation in chicken embryonic tectum.

Key words: Opitc tectum, Self-made electrode, In vivoelectroporation, Chicken embryo