Production of Transgenic Pigs Harboring the Human Erythropoietin (hEPO) Gene Using Somatic Cell Nuclear Transfer

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概要

• 論文の詳細を見る

• The production of transgenic pigs using somatic cell nuclear transfer (scNT) has been widely described, but a technique for removing nontransfected donor cells and for creating different founder animals has not yet been fully elucidated. In this study, four different expression vectors (pBC1hEPO, pMARBC1hEPO, pBC1hEPOwpre and pMARBC1hEPOwpre) were compared to determine the highest transgene expression, ideal conditions of enrichment of recombinant cells in vitro and efficiency of transgenesis following transfection into HC11 mammary epithelial cells. The highest protein expression in HC11 cells was obtained from the pMARBC1hEPOwpre expression vector. Next, we evaluated the efficiency of transgenic pig production by using geneticin (G418) selection alone or by using real-time PCR selection following G418 selection. Ideal enrichment of recombinant cells was obtained by a combination of real-time PCR and G418 selection; embryos reconstructed using donor cells selected by a combination of real-time PCR and G418 selection gave rise to nine piglets, all of which were transgenic. Among them, three founder transgenic pigs were established. Exogenous DNA fragments were shown to be integrated into chromosomes 1q2.4, 1p2.3 and 6q2.4, respectively, in these three pigs. However, the transgenic rate using G418 selection alone was only 33% (two of six pigs) and showed a very low efficacy compared with that of the combination of real-time PCR and G418 selection. Our results provide a valuable experimental model for applying and evaluating transgenic technology in pigs.

• 日本繁殖生物学会の論文

著者

• Bui Hong-thuy

  Department Of Animal Biotechnology College Of Animal Bioscience & Biotechnology Konkuk Universit

• CHOI Yun-Jung

  Department of Animal Biotechnology, College of Animal Bioscience and Technology, Konkuk University

• PARK ChangKyu

  Department of Animal Biotechnology, College of Animal Bioscience and Technology, Konkuk University

• KIM Jin-Hoi

  Department of Animal Biotechnology, College of Animal Bioscience and Technology, Konkuk University

• Kim Jin-hoi

  Department Of Animal Biotechnology College Of Animal Bioscience And Technology Konkuk University

• Choi Yun-jung

  Department Of Animal Biotechnology College Of Animal Bioscience And Technology Konkuk University

• Park Changkyu

  Department Of Animal Biotechnology College Of Animal Bioscience And Technology Konkuk University

• Cho Seong-keun

  Cho-a Biotechnology Research Institute Cho-a Pharmaceutical Co. Ltd.

• Hwang Kyu-chan

  Department Of Animal Biotechnology College Of Animal Bioscience And Technology Konkuk University

• Kim Jae-hwan

  Cha Stem Cell Institute Graduate School Of Life Science And Biotechnology Pochon Cha University

• Nguyen Van

  Department Of Anesthesiology University Of Virginia Health System

• Bui Hong-Thuy

  Department of Animal Biotechnology, College of Animal Bioscience and Technology, Konkuk University

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