Sequence analysis of proviral DNA of porcine endogenous retroviruses.
Among all species analyzed, the domestic pig seems to be the most appropriate organ donor for xenotransplantation. Porcine endogenous retroviruses (PERVs) are present in genomes of all pigs and are capable of infecting human cells in vitro thus posing a serious threat for xenotransplantation procedures. Despite the abundant distribution of PERVs integrated with porcine genome, the majority of PERV proviral DNA is not capable of expressing viral proteins unless seriously mutated. The aim of the study was to analyze PERV genome for mutations. The study was performed on blood samples from 146 pigs. Long-range polymerase chain reaction (Long-PCR) was performed with primer sets designed within long terminal repeats (LTRs). Long-PCR products of different molecular weights were obtained: 530 bp (33.1% of individuals), 580 bp (76.7%), 933 bp (100%), and 2900 bp (59.8%). Amplimers of 7200 bp were absent in 12.8% of individuals, indicating the lack of intact proviral DNA. Sequence analysis showed that most PERV proviral DNA was significantly mutated, thus suggesting the inability to express functional viral RNA; however, it cannot be ruled out that compensatory recombination processes could occur enabling replication of defective proviruses.
Machnik
G
G
Department of Biotechnology and Genetic Engineering, Medical University of Silesia, Sosnowiec, Poland. gmachnik@slam.katowice.pl
Sypniewski
D
D
Wydmuch
Z
Z
Cholewa
K
K
Mazurek
U
U
Wilczok
T
T
Smorag
Z
Z
Pacha
J
J
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004251:DNA Transposable Elements; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics*; D020077:Endogenous Retroviruses / Q000235:genetics*; D016678:Genome; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D016415:Sequence Alignment; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D013552:Swine / Q000821:virology*
D017931:DNA Primers; D004251:DNA Transposable Elements; D004279:DNA, Viral