JARB Journal of Animal Reproduction and Biotehnology

OPEN ACCESS pISSN: 2671-4639
eISSN: 2671-4663

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Journal of Embryo Transfer 2000; 15(2): 167-173

Published online August 31, 2000

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Green Fluorescent Protein 발현 토끼 수정란의 핵이식에 의한 복제

강태영, 윤희준, 노규진, 이항, 채영진, 이효종

경상대학교 수의과대학, 동물의학연구소

Abstract

The principal objective of this study was to clone transgenic embryos in order to improve the efficiency of transgenic animal production by the combination of microinjection and nuclear transplantation techniques. Mature female New Zealand White rabbits were superovulated by eCG and hCG treatments, fllowed by natural mating. Zygotes were collected from the oviducts at 18∼22 h after hCG injection by flushing with D-PBS containing 5% fetal calf serum(FCS). Two to three picoliters of green fluorescent protein(GFP) gene wa microinjected into male pronucleus. The foreign gene-injected zygotes were cultured in TCM-199 or RD medium containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% CO2 incubator. The morulae expressing GFP gene were selected and their blastomeres were separated for the use of nuclear donor. Following nuclear transplantation of fluorescence-positive morula stage blastomeres, 13 (21.3%) out of 61 fused oocytes developed to blastocyst stage and all of the cloned blastocysts expressed GFP. The results indicate that the screening of transgene in rabbit embryos by GFP detection could be a promisible method for the preselection of transgenic embryos. Also the cloning of preselected transgenic embryos by nuclear transplantatin could be efficiently applied to the multiple production of transgenic animals.

Keywords: GFP gene, unclear transplantation, transgene preselection, rabbit

Article

Journal of Embryo Transfer 2000; 15(2): 167-173

Published online August 31, 2000

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Green Fluorescent Protein 발현 토끼 수정란의 핵이식에 의한 복제

강태영, 윤희준, 노규진, 이항, 채영진, 이효종

구엘프대학교 생명의학과, 긴끼대학 동물번식학교실, 경상대학교 수의과대학, 동물의학연구소, 서울대학교 수의과대학 및 농생명공학부, 서울대학교 수의과대학 및 농생명공학부, 경상대학교 수의과대학, 동물의학연구소

Green Fluorescent Protein 발현 토끼 수정란의 핵이식에 의한 복제

강태영, 윤희준, 노규진, 이항, 채영진, 이효종

경상대학교 수의과대학, 동물의학연구소

Abstract

The principal objective of this study was to clone transgenic embryos in order to improve the efficiency of transgenic animal production by the combination of microinjection and nuclear transplantation techniques. Mature female New Zealand White rabbits were superovulated by eCG and hCG treatments, fllowed by natural mating. Zygotes were collected from the oviducts at 18∼22 h after hCG injection by flushing with D-PBS containing 5% fetal calf serum(FCS). Two to three picoliters of green fluorescent protein(GFP) gene wa microinjected into male pronucleus. The foreign gene-injected zygotes were cultured in TCM-199 or RD medium containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% CO2 incubator. The morulae expressing GFP gene were selected and their blastomeres were separated for the use of nuclear donor. Following nuclear transplantation of fluorescence-positive morula stage blastomeres, 13 (21.3%) out of 61 fused oocytes developed to blastocyst stage and all of the cloned blastocysts expressed GFP. The results indicate that the screening of transgene in rabbit embryos by GFP detection could be a promisible method for the preselection of transgenic embryos. Also the cloning of preselected transgenic embryos by nuclear transplantatin could be efficiently applied to the multiple production of transgenic animals.

Keywords: GFP gene, unclear transplantation, transgene preselection, rabbit