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In the past, several strategies have been used to generate transgenic birds. The most successful method has proven to be injection of lentiviral vector into the subgerminal cavity of the newly laid egg. In this study, we directly injected lentiviral vector into the blood vessel of HH13-15 quail embryos to produce transgenic chimeras. In the manipulated, hatched birds, the green fluorescent protein (GFP) gene driven by a cytomegalovirus (CMV) promoter was extensively expressed. All tissues analyzed were GFP-positive, and gonad cells from some of the manipulated embryos expressed GFP. The semen genome of 21.4% of mature male birds was determined to be GFP-positive by PCR, indicating these male birds were transgenic chimeras.

The purpose of this study was to produce quail-duck chimeras by transferring stage X blastoderm cells and to detect the distribution of donor cells in heterogeneous embryos using PCR. Four experimental groups were made by transferring different amounts of quail blastoderm cells into duck recipients. In early embryonic stages, donor cells labeled with PKH26 fluorescent dye were observed in the head, neural tube and gonads by fluorescent microscopy. A total of 194 duck recipient embryos were injected and 93 survived to hatch. The average hatching rate was 48% (93/194); the hatching rate showed a significant difference among all the groups (P < 0.05). Sixteen somatic chimeras were obtained, 10 of which had black feathers derived from the donor quail. The PCR results showed that donor cells were distributed in various tissues and organs of the phenotypic chimeras. This is the first report on producing Japanese quail-Peking duck chimeras by transferring quail blastoderm cells into the subgerminal cavity of the duck. This technique will provide a basis for the investigation of fertilization barriers in interspecies germline chimeras and will aid conservation of endangered wild birds.