Successful Transfer of Exogenously Introduced Gene (lacZ/MiwZ and lacZ&GFP/pkkv4-lacZ) for Generations in Chicken.

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  • EBARA Fumio
    Animal Resource Science Section, Division of Bioresource and Bioenvironmental Sciences, Graduate School, Kyushu University, Fukuoka 812-8581, Japan
  • FUJIHARA Noboru
    Animal Resource Science Section, Division of Bioresource and Bioenvironmental Sciences, Graduate School, Kyushu University, Fukuoka 812-8581, Japan

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Exogenous genes (trial 1: lacZ/MiwZ and trial 2 : lacZ&GFP/pkkv4-lacZ) introduced into germinal crescent region (GCR) of chicken embryos were confirmed to be successfully transferred to the gonads via primordial germ cells (PGCs) and remained for a long time after hatching. The injected DNA was also successfully migrated and incorporated into next generation (F1). The F1 chickens were raised until sexual maturation, then the female F1 chicken and normal males, the male F1 chicken and normal females were subjected to artificial insemination according to the routine methods. The collected fertilized eggs were incubated for 72h to obtain F2 generation and the F2 embryos were examined for the detection of introduced DNA. In trial 1, the embryos obtained from F1 female chicken were examined by X-gal staining method, and showed the expression of the lacZ gene. Furthermore, the presence of lacZ gene in the extracts from the embryos was also determined by polymerase chain reaction (PCR) analysis. However fertilized eggs obtained from F1 male chicken showed no expression and presence of the DNA. In trial 2, the expression of green fluorescent protein (GFP) gene in the F2 embryos was not detected under the fluorescent microscope. The embryos were, then, examined by X-gal staining method and no detection was observed. Finally, the presence of GFP gene in the embryo extracts was determined by PCR analysis, indicating the presence only in the embryos obtained from male F1 chicken. These results suggest that the exogenously introduced DNA was successfully transferred to the F2 generation of chicken.

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