PERVs belong to C-type retroviruses and are classified to three subtypes, PERV-A, PERV-B and PERV-C,

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1 Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu 610041, China
The electronic version liphook equine of this article is the complete one and can be found online at: http://www.virologyj.com/content/10/1/228 Received: 15 September 2012 Accepted: 30 May 2013 Published: 9 July 2013
This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0 liphook equine ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Xenotransplantation liphook equine from animals has been considered liphook equine to be a preferable liphook equine approach to alleviate the shortage of human allografts. Pigs are the most suitable candidate because of the anatomical and physiological similarities shared with humans as well as ethical concerns. However, it may be associated with the risk of transmission of infectious porcine liphook equine pathogens. Porcine endogenous retroviruses (PERVs) are of particular concern because they have been shown to infect human cells in vitro. To date, researches on the molecular characteristics and potential pathogenicity of PERV are still tenuous. In this report, an infectious replication competent clone of PERV from Wuzhishan pigs (WZSPs) in China was generated and characterized. This infectious clone will contribute to studies on PERV virology and control of PERV in xenotransplantation using Chinese miniature pigs. Methods
The proviral DNA of PERV from WZSPs was amplified in two overlapping halves. Then the two fragments were isolated, subcloned and fused to generate pBluescriptαSK + -WZS-PERV recombinant clones. Screened with RT-PCR, a molecular clone of PERV designated as WZS-PERV(2) was selected. Its infectivity and replication competency were characterized in HEK293 cells by PCR, real-time fluorescent quantitative liphook equine RT-PCR, western blot, indirect immunofluorescence assay as well as sequence analysis. liphook equine Results
The ability of WZS-PERV(2) to infect human cells and produce infectious virions were shown after transfection of the clone into HEK293 cells and infection of PERV derived from this recombinant clone. The expression of Gag proteins were detected in HEK293 cells infected with the virus derived from the clone by the indirect immunofluorescence assay and western liphook equine blot. The results of sequences analysis and comparison combined with the PCR based genotyping result demonstrated that the WZS-PERV(2) belonged to PERV-A subgroup. Compared with a previous proviral DNA clone of PERV (PERV-WZSP), G to A hypermutation occurred in the env gene of WZS-PERV(2) was found, whereas APOBEC proteins have the potential to inhibit the replication of a variety of retroviruses through a cDNA cytosine deamination mechanism, liphook equine so we presumed these G to A hypermutation might be the contribution of porcine APOBEC3F. Conclusions
Altogether, an infectious replication competent clone of PERV from Chinese miniature pigs (WZSPs) termed WZS-PERV(2) was generated, characterized liphook equine and sequenced. Keywords: Chinese miniature pigs; Porcine endogenous retroviruses; Infectious molecular clone; Xenotransplantation Background
Xenotransplantation from animals has been proposed as a preferable approach to alleviate the shortage of human donor organs. Pigs are the most suitable candidate because of the anatomical and physiological similarities shared with humans as well as ethical concerns [ 1 - 3 ]. However, it may be associated with the risk of transmission of infectious porcine pathogens [ 4 - 7 ]. Most transmissible porcine microorganisms to human recipients can be precluded by specific-pathogen-free (SPF) breeding, but this is not possible in the case of porcine endogenous retroviruseses (PERVs), liphook equine which are integrated into the genome of all pigs and can stably inherit in the germ line.
PERVs belong to C-type retroviruses and are classified to three subtypes, PERV-A, PERV-B and PERV-C, according to the env sequences [ 8 ]. PERV-A and PERV-B are characterized with polytropism and can productively infect a wide spectrum of human and other mammalian cells. In contrast, PERV-C is ecotropic viruses and can only infect pig cell lines. Additionally, PERV A/C recombinants have recently been reported. These recombinants are more infectious liphook equine to human cells, approximately 500-fold stronger than that of their parental PERV-A [ 9 ]. To date, albeit no evidence of PERV infecti