Characterization of Porcine Endogenous Retrovirus Clones from the NIH Miniature Pig BAC Library

Pigs have been considered as donors for xenotransplantation in the replacement of human organs and tissues. However, porcine endogenous retroviruses (PERVs) might transmit new infectious disease to humans during xenotransplantation. To investigate PERV integration sites, 45 PERV-positive BAC clones, including 12 PERV-A, 16 PERV-B, and 17 PERV-C clones, were identified from the NIH miniature pig BAC library. The analysis of 12 selected full-length sequences of PERVs, including the long terminal repeat (LTR) region, identified the expected of open reading frame length, an indicative of active PERV, in all five PERV-C clones and one of the four PERV-B clones. Premature stop codons were observed in only three PERV-A clones. Also, eleven PERV integration sites were mapped using a 5000-rad IMpRH panel. The map locations of PERV-C clones have not been reported before, thus they are novel PERV clones identified in this study. The results could provide basic information for the elimination of site-specific PERVs in selection of pigs for xenotransplantation.

[1]  Rodrigo Lopez,et al.  Clustal W and Clustal X version 2.0 , 2007, Bioinform..

[2]  A. Bader,et al.  Porcine endogenous retrovirus released by a bioartificial liver infects primary human cells , 2009, Liver international : official journal of the International Association for the Study of the Liver.

[3]  J. Madsen,et al.  Morphometric analyses to predict appropriate donor size for swine-to-human cardiac xenotransplantation. , 1999, Transplantation proceedings.

[4]  C. Rogel-Gaillard,et al.  Assignment1 of two new loci for gamma 1 porcine endogenous retroviruses (γ1 PERV) to pig chromosome bands 2q21 and 11q12 by in situ hybridization , 2002, Cytogenetic and Genome Research.

[5]  R. Kurth,et al.  Knockdown of porcine endogenous retrovirus (PERV) expression by PERV‐specific shRNA in transgenic pigs , 2008, Xenotransplantation.

[6]  Kyu-Tae Chang,et al.  Identification and molecular characterization of PERV Gamma1 long terminal repeats , 2009, Molecules and cells.

[7]  W Y Jung,et al.  Comparison of PERV genomic locations between Asian and European pigs. , 2010, Animal genetics.

[8]  T. Shigehisa,et al.  Prevalence of porcine endogenous retroviruses in domestic, minature, and genetically modified pigs in Japan. , 2008, Transplantation proceedings.

[9]  R. Weiss,et al.  Two sets of human-tropic pig retrovirus , 1997, Nature.

[10]  E. Park,et al.  Analysis of the molecular and regulatory properties of active porcine endogenous retrovirus gamma-1 long terminal repeats in kidney tissues of the NIH-Miniature pig , 2010, Molecules and cells.

[11]  M. Mendicino,et al.  Production of transgenic pigs that express porcine endogenous retrovirus small interfering RNAs , 2009, Xenotransplantation.

[12]  Ning Li,et al.  Characterization of two porcine endogenous retrovirus integration loci and variability in pigs , 2003, Immunogenetics.

[13]  M. Denaro,et al.  Identification of a Full-Length cDNA for an Endogenous Retrovirus of Miniature Swine , 1998, Journal of Virology.

[14]  H. Bergmans,et al.  Reappraisal of biosafety risks posed by PERVs in xenotransplantation , 2008, Reviews in medical virology.

[15]  R. Valdés-González,et al.  No evidence of porcine endogenous retrovirus in patients with type 1 diabetes after long‐term porcine islet xenotransplantation , 2010, Journal of medical virology.

[16]  J. Lunney,et al.  The piglet as a model for B cell and immune system development , 2008, Veterinary Immunology and Immunopathology.

[17]  G. Benson,et al.  Tandem repeats finder: a program to analyze DNA sequences. , 1999, Nucleic acids research.

[18]  E. Wolf,et al.  Distribution and expression of porcine endogenous retroviruses in multi‐transgenic pigs generated for xenotransplantation , 2008, Xenotransplantation.

[19]  J. Jeon,et al.  Investigation of Deletion Variation and Methylation Patterns in the 5' LTR of Porcine Endogenous Retroviruses , 2008 .

[20]  C. Rogel-Gaillard,et al.  Construction of a swine BAC library: application to the characterization and mapping of porcine type C endoviral elements , 1999, Cytogenetic and Genome Research.

[21]  L. Scobie,et al.  Identification of Exogenous Forms of Human-Tropic Porcine Endogenous Retrovirus in Miniature Swine , 2004, Journal of Virology.

[22]  E. Flory,et al.  Transcriptional Regulation of Porcine Endogenous Retroviruses Released from Porcine and Infected Human Cells by Heterotrimeric Protein Complex NF-Y and Impact of Immunosuppressive Drugs , 2002, Journal of Virology.

[23]  Wilson Ca,et al.  Porcine endogenous retroviruses and xenotransplantation. , 2008 .

[24]  F. Yoshimura,et al.  Sequence Analysis of Porcine Endogenous Retrovirus Long Terminal Repeats and Identification of Transcriptional Regulatory Regions , 2003, Journal of Virology.

[25]  U. Martin,et al.  Pig endogenous retroviruses and xenotransplantation , 2002, Xenotransplantation.

[26]  J. Denner Recombinant porcine endogenous retroviruses (PERV-A/C): a new risk for xenotransplantation? , 2008, Archives of Virology.

[27]  Yasuhiro Takeuchi,et al.  Infection of human cells by an endogenous retrovirus of pigs , 1997, Nature Medicine.

[28]  R. Allen,et al.  Characterizing and Mapping Porcine Endogenous Retroviruses in Westran Pigs , 2002, Journal of Virology.

[29]  Lee Alexander,et al.  IMpRH Server: an RH mapping server available on the Web , 2000, Bioinform..

[30]  김삼묘,et al.  “Bioinformatics” 특집을 내면서 , 2000 .

[31]  D. Milan,et al.  Construction of a whole-genome radiation hybrid panel for high-resolution gene mapping in pigs , 1998, Cytogenetic and Genome Research.