Production of Quail Chimera by Transfer of Early Blastodermal Cells and its Use for Transgenesis.

概要

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Dispersed blastodermal cells taken from the eggs of the wild-type plumage quail were microinjected into the blastoderms of unicubated fertilized eggs of sex-linked albino quail. The embryos thus treated were transferred to another quail egg shell beds and cultured for 52-53 hours at 37.5°C while being rocked at an angle of 90 degrees at 30 minutes intervals. Then, the embryos were transferred to the chicken egg shell beds and cultured at 37.5°C with rocking at a 30 degree angle until moved to hatcher just before hatching. Forty-one out of 184 injected embryos hatched and the chimeric plumage was observed in 6 hatchlings. The plumage chimerism was also found in 5 dead embryos. For transgenesis, dispersed blastodermal cells were co-cultured in vitro with a plasmid construct containing β-actin-lacZ hybrid gene (pMiwZ) and a cationic liposome (Lipofectin) for gene transfection, and then injected into the host blastoderm. The embryos were cultured in vitro for 52-53 hours. Ten out of 30 survived embryos exhibited lacZ expression in embryonic tissues as detected by histochemistry. The other 10 exhibited lacZ expression only in exrtaembryonic tissues, and the remaining 10 failed to show any sign of gene transfection. The technique described here should be highly applicable for the production of transgenic avian animals.
日本家禽学会の論文