Hatching of avian embryos under artificial environment and establishment of transgenic chicken

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This year, we studied the conditions for gene transfection to embryos. For this, we applied liposome method for embryos of various developmental stages. DNA and liposome complex was injected from heat to blood vessel. Embryos after 48h fertilization gave both the highest survival and transfection efficiency. In this experiment, only heart cells showed higher expression of the introduced gene.Then we improved this point by using protamine. For this hydrophobic residues were coupled to promine and liposome protamine complex was stabilized. By using this modified liposome, the area which showed the expression of introduced DNA was expanded.On the other hand, we developed the vector system to achieve oviduct spcific gene expression. For this we cloned ovalbimin genes from chicken fibroblast DNA.From 100,000 recombinant phages, we isolated a phage containing ovalbumin promotor region. Human insulin gene DNA was ligated to the promoter region to construct the expression vector.