Characterizing and mapping porcine endogenous retroviruses in Westran pigs.
Since porcine endogenous retroviruses (PERVs) can infect cultured human cells, they are a potential hazard to xenotransplantation. For this reason, endogenous retroviruses from the Westran (Westmead Hospital transplantation) inbred line of pigs were analyzed by using consensus primers for the type A and type B viruses to amplify 1.8-kb envelope gene fragments. After preliminary analysis with restriction enzymes KpnI and MboI, 31 clones were sequenced. Between types A and B, five recombinant clones were identified. Fifty-five percent of clones (17 of 31) had premature stop codons within the envelope protein-encoding region. Endogenous retroviruses in Westran pigs were physically mapped by fluorescence in situ hybridization (FISH) using PERV-A and PERV-B envelope clones as probes to identify at least 32 integration sites (19 PERV-A sites and 13 PERV-B sites). The chromosomal sites of integration in the Westran strain are quite different from those in the European Large White pig. The recombinant clones suggest that defective PERVs could become infective through recombination and further that PERVs might recombine with human endogenous retroviruses in xenotransplants.
Lee
Jun-Heon
JH
Centre for Advanced Technologies in Animal Genetics and Reproduction, Faculty of Veterinary Science, The University of Sydney, Sydney, New South Wales 2006, Australia.
Webb
Graham C
GC
Allen
Richard D M
RD
Moran
Chris
C
D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D018388:Codon, Terminator; D004279:DNA, Viral; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D020077:Endogenous Retroviruses / Q000235:genetics*; D016368:Frameshift Mutation; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013552:Swine / Q000821:virology*
D018388:Codon, Terminator; D004279:DNA, Viral; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases; C029312:GGTACC-specific type II deoxyribonucleases