BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
권호 표지

Expression of Cholesteryl Ester Transfer Protein cDNA using Recombinant Vaccinia Viruses

  • Jang, Moon-Kyoo (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Ahn, Byung-Yoon (Department of Genetic Engineering, College of Natural Resources, Korea University) ;
  • Huh, Tae-Lin (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University) ;
  • Bok, Song-Hae (Bioproducts Research Group, Genetic Engineering Research Institute, KIST) ;
  • Park, Yong-Bok (Department of Genetic Engineering, College of Natural Sciences, Kyungpook National University)
  • Received : 1994.10.24
  • Published : 1995.05.31
Download PDF
⟨ Previous Next ⟩

Abstract

cDNA for human cholesteryl ester transfer protein (CETP), a potent atherogenic plasma protein that redistributes the neutral lipids among lipoproteins, was expressed in recombinant vaccinia virus-infected cells (CV-1). Two insertion vectors regulated by different promoters were constructed. The vectors were introduced into human thymidine kinase-negative ($TK^-$) 1438 cells infected with wild-type vaccinia virus (WR strain). Recombinant viruses were selected with 5-bromodeoxyuridine (BUdR) and X-gal and identified with DNA dot blot analysis (vSC11-CETP and vTM1-CETP). The CETP cDNA insert in the recombinant vaccinia virus genome was identified by Southern blot analysis. Transcription of CETP cDNA in CV-1 cells infected with recombinant vaccinia virus was monitored by Northern blot analysis using the CETP cDNA as a probe. Positive signals were detected at 1.8 kb in cells infected with vSC11-CETP and at 2.3 kb in cells infected with vTM1-CETP. The recombinant vaccinia virus-infected CV-1 cells were shown to produce functional CETP when the culture medium was subjected to the CETP assay.

Keywords

References

  1. J. Biol. Chem. v.266 Agellon, L.B.;Walsh, A.;Hayek, T.;Moulin, P.;Jiang, X.C.;Shelanski, S.A.;Breslow, J.L.;Tall, A.R.
  2. Anal. Biochem. v.162 Chomczynski, P.;Sacchi, N.
  3. Nature v.327 Drayna, D.;Jamagin, A.S.;Mclean, J.;Henzel, W.;Kohr, W.;Fielding, C.;Lawn, R. https://doi.org/10.1038/327632a0
  4. Proc. Natl. Acad. Sci. USA v.86 Elroy-Stein, O.;Fuerst, T.R.;Moss, B. https://doi.org/10.1073/pnas.86.16.6126
  5. Proc. Natl. Acad. Sci. USA v.83 Fuerst, T.R.;Niles, E.G.;Studier, F.W.;Moss, B. https://doi.org/10.1073/pnas.83.21.8122
  6. J. Biol. Chem. v.262 Hesler, C.B.;Swenson, T.L.;Tall, A.R.
  7. Proc. Natl. Acad. Sci. USA v.84 Jamagin, A.S.;Kohr, W.;Fielding, C.J. https://doi.org/10.1073/pnas.84.7.1854
  8. DNA Cloning Vol. 2 Mackett, M.;Smith, G.L.;Moss, B.;Glover, D.M.(ed.)
  9. Korean Biochem. J. v.25 Park, Y.B.;Jeoung, N.H.;Kim, H.S.;Choi, M.S.
  10. Annu. Rev. Nutr. v.10 Quing, D.W.;Zilversmit, D.B. https://doi.org/10.1146/annurev.nu.10.070190.001125
  11. J. Viol. v.56 Rice, C.M.;Frankie, C.A.;Strauss, J.M.;Hruby, D.E.
  12. Molecular Cloning, A Laboratory Manual Sambrook, J.;Fritsch, E.F.;Maniatis, T.
  13. Proc. Natl. Acad. Sci. USA v.84 Yuen, L.;Moss, B. https://doi.org/10.1073/pnas.84.18.6417