Cloning, Sequencing, and Expression of cDNA Encoding Bovine Prion Protein

  • Kang, Sang-Gyun (Department of Infectious Disease, Theriogenology ,College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University) ;
  • Kang, Sung-Keun (Department of Theriogenology and Biotechnology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University) ;
  • Lee, Deog-Yong (Department of Infectious Disease, Theriogenology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University) ;
  • Park, Yong-Ho (Department of Infectious Disease, Microbiology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University) ;
  • Hwang, Woo-Suk (Department of Infectious Disease and Biotechnology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University) ;
  • Yoo, Han-Sang (Department of Infectious Disease, Theriogenology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University)
  • 발행 : 2004.04.01

초록

A normal prion protein (PrPc) is converted to a protease resistant isoform (PrPsc) by an apparent self-propagating activity in bovine spongiform encephalopathies (BSE), which is a neurodegenerative disease. The cDNA encoding bovine PrP open reading frame (ORP) in Korean cattle was cloned by polymerase chain reaction (PCR). The cloned cDNA had a length of 795 base pairs which coded for a protein of 264 amino acid residues with a calculated molecular mass of 28.6 kDa. Identities of 90, 90, 79 and 78% on nucleotide and 94, 94, 84, and 84% on amino acid sequence were shown to PrP genes from sheep, goat, human, and mouse, respectively. The cloned DNA was ligated into the pQE30 expression vector and transformed into E. coli M15. The PrP was expressed by induction with isopropyl-$\beta$-D-thiogalactoside (IPTG) and purified on the Ni-NTA affinity column. High specific activities of the recombinant PrP were observed in the fraction of pH 5.8 eluate and showed a molecular mass of-29 kDa on SDS-PAGE and Western blot analysis.

키워드

참고문헌

  1. Nature v.382 Transmission dynamic and epidemiology of BSE in British cattle Anderson,R.M.;C.A.Donnelly;N.M.Ferguson;M.E.J.Woolhouse;C.J.Watt;H.J.Udy;S.MaWhinney;S.P.Dunstan;M.E.J.Soutwood;J.W.Wilesmith;J.B.M.Ryan;L.J.Hoinville;J.E.Hillerton;A.R.Austin;G.A.H.Wells https://doi.org/10.1038/382779a0
  2. Cell v.46 Scrapie and cellular PrP isoforms are encoded by the same chromosomal gene Basler,K.;B.Oesch;M.Scott;D.Westaway;M.Walchi;D.E.Groth;M.P.Mckinley;S.B.Prusiner;C.Weissmann https://doi.org/10.1016/0092-8674(86)90662-8
  3. Neurology v.42 Nearly ubiquitous tissue distribution of the scrapie agent precursor protein Bendheim,P.E.;H.R.Brown;R.D.Rudelli;L.J.Scala;N.L.Goller;G.Y.Wen;R.J.Kascsak;N.R.Cashman;D.C.Bolton https://doi.org/10.1212/WNL.42.1.149
  4. J. Gen. Virol. v.69 Detection of prion protein mRNA in normal and scrapie-infected tissues and cell lines Caughey,B.;R.E.Race;B.Chesebro https://doi.org/10.1099/0022-1317-69-3-711
  5. Neurochem. Int. v.41 Expression in E. coli and purifiction of recombinant fragments of wild type and mutant human prion protein Corsaro,A.;S.Thellung;C.Russo;V.Villa;S.Arena;M.C.DAdamo;D.Paludi;D. Rossi Principe;G.Damonte;U.Benatti;A.Aceto;F.Tagliavini;G.Schettini;T.Florio https://doi.org/10.1016/S0197-0186(01)00137-1
  6. Blood v.91 Prion protein expression in human leukocyte differentiation Dodelet,V.C.;N.R.Cashman
  7. J. Vet. Med. Sci. v.59 Characterization of the bovine prion protein gene: The expression requires interaction between the promoter and intron Inoue,S.;M.Tanaka;M.Horiuchi;N.Ishiguro;M.Shinagawa https://doi.org/10.1292/jvms.59.175
  8. Clin. Microbiol. Rev. v.12 Cellular biology of prion disease Harris,D.A.
  9. Proc. Natl. Acad. Sci. USA v.90 Location of the mRNA for a chicken prion protein by in situ hybridization Harris,D.A.;P.Lele;W.D.Snider https://doi.org/10.1073/pnas.90.9.4309
  10. Structure v.7 Prion protein interconversions and the transmissible spongiform encephalopathies Horiuchi,M.;B.Caughey https://doi.org/10.1016/S0969-2126(00)80049-0
  11. J. Microbiol. Biotechnol. v.11 Effects of environmental factors on in vivo folding of Bacillus macerans cyclodextrin glycosyltransferase in recombinant Escherichia coli Jin,H.H.;N.S.Han;D.H.Kweon;Y.C.Park;J.H.Seo
  12. J. Microbiol. Biotechnol. v.10 Refolding of Bacillus macernas cyclodextrin glucanotransferase expressed as inclusion bodies in recombinant Escherichia coli Kim,C.I.;M.D.Kim;Y.C.Park;N.S.Han;J.H.Seo
  13. Proc. Natl. Acad. Sci. USA v.83 Molecular cloning and complete sequence of prion protein cDNA from mouse brain infected with the scrapie agent Locht,C.;B.Chesebro;R.Race;J.M.Keith https://doi.org/10.1073/pnas.83.17.6372
  14. Development v.115 The prion protein gene: A role in mouse embryogenesis? Manson,J.;J.D.West;V.Thomson;P.McBride;M.H.Kaufman;J.Hope
  15. Neuron v.14 Developmental expression of the prion protein gene in glial cells Moster,M.;R.J.Colello;U.Pott;B.Oesch https://doi.org/10.1016/0896-6273(95)90307-0
  16. J. Comp. Pathol. v.113 Spongiform central nervous system myelinopathy in African dwarf goats Obermaier,G.;H.A.Kretzschmar;A.Hafner;D.Heubeck;E.Dahme https://doi.org/10.1016/S0021-9975(05)80121-5
  17. J. Microbiol. Biotechnol. v.12 Expression and purification of an ACE-inhibitory peptide multimer from synthetic DNA in Escherichia coli Oh,K.S.;Y.S.Park;H.C.Sung
  18. J. Gen. Virol. v.75 SCID mouse spleen does not support scrapie agent relication O'Rourke,K.I.;T.T.Huff;C.W.Leathers;M.M.Robinson;J.R.Gorham https://doi.org/10.1099/0022-1317-75-6-1511
  19. Proc. Natl. Acad. Sci. USA v.90 Conversion of alpha helices into beta-sheet features in the formation of the scrapie prion protein Pan,K.M.;M.Baldwin;J.Nguyen;M.Gasset;A.Sebran;D.Groth;I.Mehlborn;Z.Huang;R.J.Fletterick;F.E.Cohen;B.S.Prusiner https://doi.org/10.1073/pnas.90.23.10962
  20. Anal. Biochem. v.236 Semipreparative chromatographic method to purify the normal cellular isoform of the prion protein in nondenatured form Pergami,P.;H.Jaffe;J.Safar https://doi.org/10.1006/abio.1996.0132
  21. Science v.216 Novel proteinaceous infectious particles cause scrapie Prusiner,S.B. https://doi.org/10.1126/science.6801762
  22. Science v.278 Prion disease and the BSE crisis Prusiner,S.B. https://doi.org/10.1126/science.278.5336.245
  23. J. Infect. Dis. v.167 Immunologic and molecular bioloc studies of prion proteins in bovine spongiform encephalopathy Prusiner,S.B.;M.Fuzi;M.Scott;D.Serban;H.Serban;A.Taraboulos;J.M.Gabriel;G.A.Wells;J.W.Wilesmith;R.Bradley https://doi.org/10.1093/infdis/167.3.602
  24. Proc. Natl. Acad. Sci. USA v.99 Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences Strausberg,R.L.;E.A.Feingold;L.H.Grouse;J.G.Derge;R.D.Klausner;F.S.Collins;L.Wagner;C.M.Shenmen;G.D.Schuler;S.F.Altschul;B.Zeeberg;K.H.Buetow;C.F.Schaefer;N.K.Bhat;R.F.Hopkins;H.Jordan;T.Moore;S.I.Max;J.Wang;F.Hsieh;L.Diatchenko;K.Marusina;A.A.Farmer;G.M.Rubin;L.Hong;M.Stapleton;M.B.Soares;M.F.Bonaldo;T.L.Casavant;T.E.Scheetz;M.J.Brownstein;T.B.Usdin;S.Toshiyuki;P.Carninci;C.Prange;S.S.Raha;N.A.Loquellano;G.J.Peters;R.D.Abramson;S.J.Mullahy;S.A.Bosak;P.J.McEwan;K.J.McKernan;J.A.Malek:P.H.Gunaratne;S.Richards;K.C.Worley;S.Hale;A.M.Garcia;L.J.Gay;S.W.Hulyk;D.K.Villalon;D.M.Muzny;E.J.Sodergren;A.Madan;S.Rodrigues;A.Sanchez;M.Whiting;A.Madan;A.C.Young;Y.Shevchenko;G.G.Bouffard;R.W.Blakesley;J.W.Touchman;E.D.Green;M.C.Dickson;A.C.Rodriguez;J.Grimwood;J.Schmutz;R.M.Myers;Y.S.N.Butterfield;M.I.Krzywinski;U.Skalska;D.E.Smailus;A.Schnerch;J.E.Schein;S.J.M.Jones;M.A.Marra;Mammalian Gene Collectin Program Team https://doi.org/10.1073/pnas.242603899
  25. Arch.Virol. v.146 PrP genotype in Sarda breed sheep and its relevance to scrapie Vaccari,G.;R.Petraroli;U.Agrimi;C.Eleni;M.G.Perfetti;M. A. Di Bari;L.Morelli;C.Ligios;L.Busani;R.Nonno;G. Di Guardo https://doi.org/10.1007/s007050170050
  26. Eur. J. Biochem. v.251 Large-scale production, purification and refolding of the full-length cellular prion protein from Syrian golden hamster in E. coli using the glutathione S-transferase-fusion system Volkel,D.;W.Blankenfeldt;D.Schomburg https://doi.org/10.1046/j.1432-1327.1998.2510462.x
  27. FEBS Lett. v.389 The ninth data lecture. Molecular biology of transmissible spongiform encephalopathies Weissmann,C. https://doi.org/10.1016/0014-5793(96)00610-2
  28. Curr. Opin. Neurobiol. v.7 Boving spongiform encephalopathy and early onset variant Creutzfeldt-Jakob disease Weissmann,C.;A.Aguzzi https://doi.org/10.1016/S0959-4388(97)80091-8
  29. Virus Genes v.6 Comparative sequence analysis and expression of bovine PrP gene in mouse L-929 cells Yoshimoto,J.;T.Iinuma;N.Ishiguro;M.Horiuchi;M.Imamura;M.Shinagawa https://doi.org/10.1007/BF01703083