Two pig endogenous retroviruses (PERV), PERV-A and -B, productively infect human cells and are therefore considered to constitute a potential risk in pig-to-human xenotransplantation. A PCR-based cloning technique to isolate infectious PERV proviruses was established. Overlapping 3' half and 5' halves of PERV proviral genomes were amplified using DNA extracted from human 293 cells infected with PERV-A or -B. These clones were fused at a unique restriction site in the overlapping region and tested for their infectivity. Representative constructs possessed the same infectious properties as their parent isolates. We also developed a polyclonal anti-PERV serum by using recombinant PERV capsid protein derived from one of the infectious constructs as immunogen and established an immunocytological method for detection and titration of PERV infection. This detection method proved to be more sensitive than the current method of choice (transfer of MLV-lacZ vectors) for infectivity assessment of PERV. These findings should be considered for future characterization of PERV isolates.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1099/0022-1317-83-9-2231 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Molecular mechanisms impact on fluoroquinolone resistance among E.coli from enteric carriage monitoring before prostate biopsy and earliest description of qnrB81.
Sci Rep
November 2024
Faculty of Medicine of Tunis - LR99ES09 Research Laboratory «Antimicrobial resistance», University of Tunis El Manar, 1007, Tunis, Tunisia.
Article Synopsis
- Fluoroquinolone-resistant (FQs-R) E. coli from patients undergoing prostate biopsies are increasingly common, raising concerns about using FQs for infection prevention.
- A study between 2016 and 2018 found that 61.06% of patients carried FQs-R Enterobacterales, primarily E. coli, with varying resistance profiles linked to specific genetic mutations.
- The research highlighted that these resistance mechanisms, including mutations in the gyrA and parC genes as well as certain plasmid-mediated resistance genes, significantly elevate the Minimum Inhibitory Concentrations (MICs), posing a risk for post-procedure infections.
An improved DNA extraction method in okra for rapid PCR detection of Okra enation leaf curl virus from diverse Indian regions.
Arch Microbiol
November 2024
Plant Molecular Virology Laboratory, Department of Genetics and Plant Breeding, Chaudhary Charan Singh University, Meerut, Uttar Pradesh, 250004, India.
The extraction of DNA from okra (Abelmoschus esculentus) is challenging due to its high mucilage and polysaccharide content, which can hinder both the yield and quality of DNA. In this study, an improved DNA isolation method is described incorporating a key modification being the use of solution I (1 M NaCl and 2% Sarcosyl) as a pre-treatment before applying the CTAB buffer, resulting in high-purity genomic DNA in just 1 h and 45 min., making it suitable for handling large sample sizes due to its rapid processing capabilities.
View Article and Find Full Text PDFDevelopment of a lambda Red based system for gene deletion in Chlamydia.
PLoS One
November 2024
Department of Medicine, University of Washington, Seattle, Washington, United States of America.
Article Synopsis
- * Researchers successfully deleted the incA gene in C. trachomatis and verified the gene's absence through genetic testing and microscopy, observing changes in cell behavior.
- * The system also successfully deleted multiple virulence factor genes in different Chlamydia strains, showing promise for advancing genetic manipulation techniques in this bacterium.
Expression Cloning of Antibodies from Single Human B Cells.
Methods Mol Biol
October 2024
Division of B Cell Immunology, German Cancer Research Center (DKFZ), Heidelberg, Germany.
The majority of lymphomas originate from B cells at the germinal center stage. Preferential selection of B-cell clones by a limited set of antigens has been suggested to drive lymphoma development. While recent studies in B-cell chronic lymphocytic leukemia (CLL) have shown that self-reactive B-cell receptors (BCR) can generate cell-autonomous signaling and proliferation, our knowledge about the role of BCRs for the development or survival of other lymphomas remains limited.
View Article and Find Full Text PDFPCR GeneScan Analysis of Rearranged Immunoglobulin or T-Cell Receptor Genes for Clonality Diagnostics in Suspect Lymphoproliferations.
Methods Mol Biol
October 2024
Department of Immunology, Laboratory Medical Immunology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
Assessment of the presence of clonal lymphoproliferations via polymerase chain reaction (PCR)-based analysis of rearranged immunoglobulin (IG) or T-cell receptor (TR) genes is a valuable method in the diagnosis of suspect lymphoproliferative disorders. Additionally, this methodology can be used for evaluating dissemination of lymphoma cells and for studying the clonal relationship between multiple (different locations) and consecutive (over time) lymphomas. Here we describe an integrated approach to assess clonality via analysis of Ig heavy chain (IGH), Ig kappa (IGK), TCR beta (TRB), and TCR gamma (TRG) gene rearrangements, based on the standardized multiplex PCRs as originally developed by the European BIOMED-2 consortium (currently named EuroClonality).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!