, Volume 55, Issue 4, pp 262–270

Characterization of two porcine endogenous retrovirus integration loci and variability in pigs

  • Marie Gorbovitskaia
  • Zhaoliang Liu
  • Noelle Bourgeaux
  • Ning Li
  • Zhengxing Lian
  • Patrick Chardon
  • Claire Rogel-Gaillard
Original Paper


The pig (Sus scrofa) is a potential organ donor for man but porcine endogenous retroviruses (PERVs) represent an important concern for patients, and identification or engineering of PERV-free pigs suitable for xenotransplantation is a major undertaking. Consequently, studies of variability in pigs for the presence of PERVs at specific loci are a prerequisite. We identified genomic flanking sequences of two PERVs cloned in bacterial artificial chromosomes, a replication-competent PERV-A at locus 1q2.4 and a defective PERV-B at locus 7p1.1–2. PERV-A is embedded in the second repeat of a tandem of eight 190 bp repeats. A short duplicated 4 bp cellular motif, AGAC, was found at each flank of PERV-A and a degenerate 4 bp motif was found for PERV-B. At each locus, the PERV flanks matched expressed sequence tags available in public databases. Primer pairs were designed to amplify either genomic flanks or PERV-genomic junctions. Polymerase chain reaction screening was performed on pigs from 11 distinct Chinese breeds and from the European Large White breed. PERV-B at locus 7p1.1–2 was detected in all animals whereas the presence of PERV-A at locus 1q2.4 was variable. Our results suggest that a genetic selection can be designed to identify animals lacking a potentially active PERV at a specific locus and that Chinese and European pig breeds represent large biodiversity reservoirs to explore. Our results point also to the existence of PERVs that might be fixed in the pig genome, and that might not be eliminated by classical genetic selection.


Xenotransplantation Porcine endogenous retrovirus Genetic variability Pig Proviral integration 


  1. Akiyoshi DE, Denaro M, Zhu H, Greenstein JL, Banerjee P, Fishman JA (1998) Identification of a full-length cDNA for an endogenous retrovirus of miniature swine. J Virol 72:4503–4507PubMedGoogle Scholar
  2. Blusch JH, Seelmeir S, von der Helm K (2002) Molecular and enzymatic characterization of the porcine endogenous retrovirus protease. J Virol 76:7913–7917CrossRefPubMedGoogle Scholar
  3. Clémenceau B, Jegou D, Martignat L, Sai P (2002) PERV infection of mouse and human cells by SFP pig islets in nude mice. Diabetologia 45:914–923CrossRefGoogle Scholar
  4. Coffin JM (1996) Retroviridae; the viruses and their replication. In: Fields BN, Knipe DM, Howley PM (eds) Fields Virology, 3rd edn. Lippincott-Raven, Philadelphia, Md, pp 1767–1847Google Scholar
  5. Czauderna F, Fisher N, Boller K, Kurth R, Tönjes RR (2000) Establishment and characterization of molecular clones of porcine endogenous retroviruses replicating on human cells. J Virol 74:4028–4038CrossRefPubMedGoogle Scholar
  6. Deng YM, Tuch BE, Rawlinson WD (2000a) Transmission of porcine endogenous retroviruses in severe combined immunodeficient mice xenotransplanted with fetal porcine pancreatic cells. Transplantation 70:1010–1016PubMedGoogle Scholar
  7. Deng, YM, Lee JH, Moran C, Jin JH, Tuch BE, Rawlinson WD (2000b) Mapping dispersed repetitive loci using semi-specific PCR cloning and somatic cell hybrid mapping. Nucleic Acids Res 28:E103PubMedGoogle Scholar
  8. Heneine W, Tibell A, Switzer WM, Sandstrom P, Vazquez Rosales G, Mathews A, Korsgren O, Chapman LE, Folks TM, Groth CG (1998) No evidence of infection with porcine endogenous retrovirus in recipients of porcine islet-cell xenografts. Lancet 352:695–699PubMedGoogle Scholar
  9. Herring C, Quinn G, Bower R, Parsons N, Logan NA, Brawley A, Elsome K, Whittam A, Fernandez-Suarez XM, Cunningham D, Onions D, Langford G, Scorbie L (2001) Mapping full-length porcine endogenous retroviruses in a Large White pig. J Virol 75:12252–12265CrossRefPubMedGoogle Scholar
  10. Hughes SH, Mutschler A, Bishop JM, Varmus HE (1981) A Rous sarcoma virus provirus is flanked by short direct repeats of a cellular DNA sequence present in only one copy prior to integration. Proc Natl Acad Sci U S A 78:4299–4303Google Scholar
  11. Krach U, Fischer N, Czauderna F, Tönjes RR (2001) Comparison of replication-competent molecular clones of porcine endogenous retrovirus class A and class B derived from pig and human cells. J Virol 75:5465–5472CrossRefPubMedGoogle Scholar
  12. Lai L, Kolber-Simonds D, Park KW, Cheong HT, Greenstein JL, Im GS, Samuel M, Bonk A, Rieke A, Day BN, Murphy CN, Carter DB, Hawley RJ, Prather RS (2002) Production of alpha-1,3-galactosyltransferase knockout pigs by nuclear transfer cloning. Science 295:1089–1092CrossRefPubMedGoogle Scholar
  13. Le Tissier P, Stoye JP, Takeuchi Y, Patience C, Weiss RA (1997) Two sets of human-tropic pig retrovirus. Nature 389:681–682PubMedGoogle Scholar
  14. Lee JH, Webb GC, Allen RDM, Moran C (2002) Characterizing and mapping porcine endogenous retroviruses in Westran pigs. J Virol 76:5548–5556CrossRefPubMedGoogle Scholar
  15. Martin U, Steinhoff G, Kiessig V, Chikobava M, Anssar M, Morschheuser T, Lapin B, Haverich A (1998) Porcine endogenous retrovirus (PERV) was not transmitted from transplanted porcine endothelial cells to baboons in vivo. Transplant Int 11:247–251CrossRefGoogle Scholar
  16. Martin U, Steinhoff G, Kiessig V, Chikobava M, Anssar M, Morschheuser T, Lapin B, Haverich A (1999) Porcine endogenous retrovirus is transmitted neither in vivo nor in vitro from porcine endothelial cells to baboons. Transplant Proc 31:913–914CrossRefPubMedGoogle Scholar
  17. Niebert M, Rogel-Gaillard C, Chardon P, Tönjes RR (2002) Characterization of chromosomally assigned replication-competent gamma porcine endogenous retroviruses derived from a Large White pig and expression in human cells. J Virol 76:2714–2720CrossRefPubMedGoogle Scholar
  18. Nikbakht KN, Boone LR, Glover PL, Myer FE, Yang WK (1987) Characterization of a molecular clone of RFM/Un mouse chromosomal DNA that contains a full-length endogenous murine leukaemia virus-related proviral genome. J Gen Virol 68:683–693PubMedGoogle Scholar
  19. Oldmixon BA, Wood JC, Ericsson TA, Wilson CA, White-Scharf ME, Andersson G, Greenstein JL, Schuurman HJ, Patience C (2002) Porcine endogenous retrovirus transmission characteristics of an inbred herd of miniature swine. J Virol 76:3045–3048CrossRefPubMedGoogle Scholar
  20. Paradis K, Langford G, Long Z, Heneine W, Sandstrom P, Switzer WM, Chapman LE, Lockey C, Onions D, the XEN 111 Study Group, Otto E (1999) Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue. Science 285:1236–1241PubMedGoogle Scholar
  21. Patience C, Takeuchi Y, Weiss RA (1997) Infection of human cells by an endogenous retrovirus of pigs. Nat Med 3:282–286PubMedGoogle Scholar
  22. Patience C, Patton GS, Takeuchi Y, Weiss RA, McClure MO, Rydberg L, Breimer ME (1998) No evidence of pig DNA or retroviral infection in patients with short-term extracorporeal connection to pig kidneys. Lancet 352:699–701PubMedGoogle Scholar
  23. Patience C, Switzer WM, Takeuchi Y, Griffiths DJ, Goward ME, Heneine W, Stoye JP, Weiss RA (2001) Multiple groups of novel retroviral genomes in pigs and related species. J Virol 75:2771–2775CrossRefPubMedGoogle Scholar
  24. Qari SH, Magre S, Garcia-Lerma JG, Hussain AI, Takeuchi Y, Patience C, Weiss RA, Heneine W (2001) Susceptibility of the porcine endogenous retrovirus to reverse transcriptase and protease inhibitors. J Virol 75:1048–1053CrossRefPubMedGoogle Scholar
  25. Rogel-Gaillard C, Bourgeaux N, Billault A, Vaiman M, Chardon P (1999) Construction of a swine BAC library: application to the characterization and mapping of porcine type C endoviral elements. Cytogenet Cell Genet 85:205–211CrossRefPubMedGoogle Scholar
  26. Rogel-Gaillard C, Hayes H, Bourgeaux N, Chardon P (2001) Assignment of two new loci for gamma 1 porcine endogenous retroviruses (γ1 PERV) to pig chromosome bands 2q21 and 11q12 by in situ hybridization. Cytogenet Cell Genet 95:112–113CrossRefPubMedGoogle Scholar
  27. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar
  28. Scherdin U, Rhodes K, Breindl M (1990) Transcriptionally active genome regions are preferred targets for retroviral integration. J Virol 64:907–912PubMedGoogle Scholar
  29. Shih CC, Stoye JP, Coffin JM (1988) Highly preferred targets for retrovirus integration. Cell 53:531–537PubMedGoogle Scholar
  30. Takeuchi Y, Patience C, Magre S, Weiss RA, Banerjee PT, Le Tissier P, Stoye JP (1998) Host range and interference studies of three classes of pig endogenous retrovirus. J Virol 72:9986–9991PubMedGoogle Scholar
  31. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTALW: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedGoogle Scholar
  32. Tristem M, Kabat P, Lieberman L, Linde S, Karpas A, Hill F (1996) Characterization of a novel murine leukemia virus-related subgroup within mammals. J Virol 70:8241–8246PubMedGoogle Scholar
  33. van der Laan LJW, Lockey C, Griffeth BC, Frasier FS, Wilson CA, Onions DE, Hering BJ, Long Z, Otto E, Torbett BE, Salomon DR (2000) Infection by porcine endogenous retrovirus after islet xenotransplantation in SCID mice. Nature 407:90–94PubMedGoogle Scholar
  34. Wilson CA, Wong S, Muller J, Davidson CE, Rose TM, Burd P (1998) Type C retrovirus released from porcine primary peripheral blood mononuclear cells infects human cells. J Virol 72:3082–3087PubMedGoogle Scholar
  35. Wilson CA, Wong S, VanBrocklin M, Federspiel MJ (2000) Extended analysis of the in vitro tropism of porcine endogenous retrovirus. J Virol 74:49–56PubMedGoogle Scholar
  36. Weiss RA (1998) Transgenic pigs and virus adaptation. Nature 391:327–328PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Marie Gorbovitskaia
    • 1
  • Zhaoliang Liu
    • 2
  • Noelle Bourgeaux
    • 1
  • Ning Li
    • 2
  • Zhengxing Lian
    • 2
  • Patrick Chardon
    • 1
  • Claire Rogel-Gaillard
    • 1
  1. 1.Laboratoire de Radiobiologie et Etude du GénomeUMR INRA CEA 13.314 Jouy-en-JosasFrance
  2. 2.The State Key Laboratory of Agro-biotechnologyChina Agricultural UniversityBeijingChina

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