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Analysis of Transgene Intergration Efficiency into Porcine Fetal Fibroblast using Different Transfection Methods
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  • Analysis of Transgene Intergration Efficiency into Porcine Fetal Fibroblast using Different Transfection Methods
  • Analysis of Transgene Intergration Efficiency into Porcine Fetal Fibroblast using Different Transfection Methods
저자명
Kim. Baek-Chul,Kim. Hong-Rye,Kim. Myung-Yoon,Park. Chang-Sik,Jin. Dong-Il
간행물명
Reproductive & developmental biology
권/호정보
2009년|33권 2호|pp.113-117 (5 pages)
발행정보
한국동물번식학회
파일정보
정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Animals produced by somatic cell nuclear transfer (SCNT) using genetically modified cells are almost always transgenic, implying that this method is more efficient than the traditional pronuclear microinjection method. Most somatic cells for SCNT in animals are fetus-derived primary cells and successful gene integration in somatic cells will depend on transfection condition. The objective of this study is to evaluate the efficiency of electroporation (Microporator) and liposome reagents (F-6, F-HD, W-EX, W-Q, W-M) for tissue-type plasminogen activator (tPA) gene transfection and to estimate the overall efficiency of transfection of Korean native pig fetal fibroblast cells (KNPFF). Electroporation showed significantly higher transfection efficiency than liposome reagents with regard to the transfection of in vitro cultures in the early stages of development (41.7% with Microporator vs. 18.3% with F-6, 20.0% with F-HD 18.5% with W-EX, 5.0% with W-M and 6.3% W-Q,). Colonies identified as tPA-positives were treated once more with G418 for 10 to 14 days and growing colonies were selected again. When the cells of newly selected colonies were subjected to single-cell PCR, reselection of colonies following second round of G418 selection increased the rate of transgene integration per each colony. These results suggest that transfection with electroporation is the most efficient and the second rounds of G418 selection may be an effective method for transfection of porcine fetal fibroblast cells.