Introduction of Exogenous Genes into Chicken Embryos by Electroporation Using a Needle Type Electrode.

Abstract

In this experiment, an exogenous gene, green fluorescent protein (GFP), was introduced into chicken blastodermal cells (stage X) just after oviposition using two types of electrode, parallel and needle, comparing embryonic survivability and subsequent development of embryos. The needle type electrode was prepared with 27-gauge needle of syringe which was covered with an insulating paint to isolate electricity. Electricity employed in this study was 10v (needle type) and 200v (parallel type) of pressure, 10msec of pulse and 10 times of repeated intervals. The embryonic survivabilities were 65% for needle type and 53% for parallel one, respectively, though no significant (P>0.05) differ-ence was observed between two types of electrode. The expression rates of GFP in embryonic tissues were 48% for needle type and 7% for parallel one, respectively, showing significant (P<0.01) difference between two types of electrode. In all of these treated embryos, GFP expression was mosaic manner for both types of electrode, indicating especially positive expression in embryonic tissues for needle type and in vitelline membrane for parallel one. Therefore, the present results suggest that the needle type of electrode might be superior to parallel one for introducing foreign genes into chicken embryos when used for electroporation.