Genetic variation of BIV isolates characterized by PCR using degenerate primers

  • Kwon, Oh-Sik (Department of Biology, Keimyung University) ;
  • Sninsky, John J. (Roche Molecular System Inc.)
  • Published : 1995.09.01

Abstract

The PCR was employed to detect and characterize the bovine immunodeficiency-like virus (BIV), which is a newly recognized member of the I entivirinae of the retroviruses. Degenerate primers representing the conserved regions in the pol genes of the Lentivirinae, were used to detect proviral DNA obtained from the bovine embryonic spleen cell cultures infected with BIV. The PCR amplified DNA fragment was molecularly cloned and sequenced. The BIV DNA fragment contained a sequence identical to that reported by Garvey et al. (Garvey et al., 1990. Virology, 175, 391-409). With the degenerate primers, peripheral blood mononuclear cells (PBMCs) of sick cattle and cells cultured with BIV were tested to determine genetic variation of BIV pol conserved sequence. We found the sequence heterogeneity within cultures and most variations occurred at the third base of codons that would not lead to amino acid substitutions. Another change was GAG (Glu) to AAG (Lys) within the BIV isolates. Interestingly, the altered sequence is also found in other lentiviruses such as HIV-2, SIV mac, CAEV and EIAV.

Keywords

References

  1. Vet. Micro. v.20 Serological detection of multiple retroviral infection in cattle: Bovine leukemia virus, bovine syncytial virus and bovine visna virus Amborski,G.F.;J.L.Lo;C.L.Seger
  2. J. Virol. v.12 Ultrastructural studies of a visna-like syncytia-producing virus from cattle with lymphocytosis Boothe,A.D.;M.J.Van Der Maaten
  3. Science v.223 Major pol gene progenitors in the evolution of oncoviruses Chiu,I.M.;R.Callahan;S.R.Tronick;J.Schom;S.A.Aaronson
  4. Nature v.317 Nucleotide sequence evidence for relationship of AIDS retrovirus to lentiviruses Chiu,I.M.;A.Yaniv;J.E.Dahlberg;A.Gazit;S.F.Skuntz;S.R.Tronick;S.A.Aaronson
  5. Vet. Micro. v.31 Seroprevalence of bovine immunodeficiency-like virus and bovine leukemia virus in a dairy cattle herd Cockerell,G.L.;.;W.A.Jensen;J.Rovnak;W.H.Ennis;M.A.Gonda
  6. Virology v.175 Nucleotide sequence and genome organization of biologically active proviruses of the bovine immunodeficiency-like virus Garvey,K.J.;M.S.Oberste;J.E.Elser;M.J.Braun;M.A.Gonda
  7. Nature v.330 Characterization and molecular cloning of a bovine lentivirus related to the human immunodeficiency virus Gonda,M.A.;M.J.Braun;S.G.Carter;T.A.Kost;J.W.Bess Jr.;L.O.Arthur;M.J. Van Der Maaten
  8. HIV detection by genetic engineering methods Genetic variation among isolates of HIV-1 and HIV-2 Hahn,B.H.;G.M.Shaw;P.A.Luciw(ed.);K.S.Steimer(ed.)
  9. J. Inst. Nat. Sci. v.10 A study of polymerase chain reaction using degenerate primers for lentivirus detection Kwon,O.S.
  10. Mol. Cells v.4 Diagnosis of bovine immunodeficiency-like virus infection by optimizing polymerase chain reaction Kwon,O.S.;G.H.Theilen
  11. Nature v.327 Site-specific mutagenesis of AIDS virus reverse transcriptase Larder,B.A.;D.J.M.Purifoy;K.L.Powell;G.Darby
  12. PCR protocols: a guide to methods and application Novel viruses Mack,D.;O.S.Kwon;F.Faloona;M.A.Innis(ed.);D.H.Gelfand(ed.);J.J.Sninsky(ed.);T.J.White(ed.)
  13. Proc. Natl. Acad. Sci. USA v.85 A sensitive method for the identification of uncharacterized viruses related to known virus group: Hepadnavirus model system Mack,D.;J.J.Sninsky
  14. Methods Enzymol. v.155 Specific synthesis of DNA in vitro via polymerase catalyzed chain reaction Mullis,K.B.;F.Faloona
  15. Science v.242 Fidelity of HIV-1 reverse transcriptase Preston,B.D.;B.J.Poiesz;L.A.Loeb
  16. Science v.242 The accuracy of reverse transcriptase from HIV-1 Roberts,J.D.;K.Bebenek;T.A.Kunkel
  17. Science v.239 Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase Saiki,R.K.;D.H.Gelfand;S.Stoffel;S.J.Scharf;R.Higuchi;G.T.Horn;K.B.Mullis;H.A.Erlich
  18. Molecular cloning: A laboratory manual Sambrook,J.;E.F.Fritsch;T.Maniatis
  19. Science v.233 Direct cloning and sequence analysis of enzymatically amplified genomic sequence Scharf,S.J.;G.T.Horn;H.A.Erlich
  20. J. Virol. v.67 Isolation and characterization of new wild-type isolates of bovine lentivirus Suarez,D.L.;M.J.Van Der Maaten;C.Wood;C.A.Whetstone
  21. J. Natl. Cancer v.49 Isolation of a virus from cattle with persistent lymphocytosis Van Der Maaten;M.J.;A.D.Boothe;C.L.Seger
  22. LAV. Cell v.40 Nucleotide sequence of AIDS virus Wain-Hobson,S.;P.Sonigo;O.Danos;M.Alizon
  23. Arch. Virol. v.116 A western blot assay for the detection of antibodies to bovine immunodeficiency-like virus in experimentally inoculated cattle, sheep and goats Whetstone,C.A.;M.J.Van Der Maaten;J.M.Miller