• Title/Summary/Keyword: human HepG2 cells

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Construction of Retrovirus Vector System for the Regulation of Recombinant hTPO Gene Expression (재조합 hTPO 유전자의 발현 조절을 위한 Retrovirus Vector System의 구축)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Do-Hyang;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.161-167
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    • 2007
  • In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.