닭 초기 배아의 유전자 미세주입과 유전자 발현에 관한 연구 = Microinjection and Expression of Marker Gene in the Early Chicken Embryo

다국어 초록 (Multilingual Abstract)

This study was carried out to examine the expression of marker genes and integration of plasmid DNA into the germ cells in young chicken embryos. The RSVLTR/βG2 plasmid contains the lacZ gene under the control of rous sarcoma virus (RSV) long terminal repeat(LTR) promoter. After a square window of 5㎜ per side was cut in the side of the egg with a dentistry drill, the transfection cocktail of calcium-phosphate or lipofectin with plasmid DNA was microinjected in the area of blastoderm and germinal crescent whose developmental stages were stage X and stage 6 to 8, respectively. Microinjected eggs were sealed, and the eggs were returned to the incubator with the $quot;window$quot; side up overnight. Microinjected embryos with plasmid DNA were screened with Xgal, and the marker gene was expressed in the brain, notochord and other parts of body of 1.5∼4.5 day old embryos, suggesting that developing stem cells in unincubated blastoderms or 1∼4 somite embryos can be transfected with plasmid vectors. The possibility of germ line integration with plasmid DNA by direct microinjection into early chicken embryos was determined. Transfection of stem cells for gonads in the blastoderm or germinal crescent with plasmid vectors was observed. Positive primordial germ cells in the gonad were not observed by plasmid DNA microinjection into unincubated or 24hr incubated embryos in this study. However, the expression of plasmid DNA with RSVLTR promoter in the early chicken embryonic cell shows the possibility of transgenic chicken production by direct microinjection with plasmid DNA. Also genetic manipulation of chicken production traits such as disease resistance, growth and production may he possible in the future.