No abstract

By using in ovo electroporation, the firefly luciferase gene was transfected to living chicken embryos at 2 days of incubation.At 48hrs after transfection, expression of the firefly luciferase gene was monitored in ovo by using a single photon imaging counter.Strong bioluminescence detected from the incubating embryo suggests that in ovo electroporation would provide a useful and efficient nonviral means of foreign gene transfection to somatic cells of living chicken embryos.

The experiments were done to investigate the effects of incubation time and injection sites on the expression of a reporter gene transfected by lipofection to somatic cells of chicken embryos in ovo. Gene expression was detected by X-gal histochemical staining at 48hrs after transfection.The results showed that when the bacterial lacZ gene was transfected to the embryonic body at 24 or 48hrs of incubation, no difference in the gene expression was found between the incubation periods, but the viability of embryos was higher when injected at 48hrs than 24hrs of incubation.No effect of injection sites on gene transfection efficiency was found in the bacterial lacZ gene expression, although the viability was higher by injecting with the reporter gene in the centre than in the rest of the embryonic body. It was concluded, therefore, that for in ovo lipofection method, 48-hr incubated embryos should be used, and DNA be injected in the centre of embryonic body not because of the enhanced gene expression but because of the improved viability.

No abstract