DNA Amplification Using Promega's PowerPlex Fusion Systems (5C and 6C).

Multiplex amplification and typing of short tandem repeat (STR) loci enable the rapid characterization of forensic samples for human identification purposes with the potential for high discriminatory power. Many commercial multiplex kits are available for consideration and purchase by DNA testing laboratories, including the PowerPlex Fusion 5C and PowerPlex Fusion 6C Systems offered by Promega Corporation. Herein we describe full-volume and half-volume amplification protocols utilizing extracted DNA for these respective multiplex kits, as well as procedures for direct amplification of lytic and nonlytic storage card punches and swabs.

Deletion of the Chemokine Binding Protein Gene from the Parapoxvirus Orf Virus Reduces Virulence and Pathogenesis in Sheep.

Orf virus (ORFV) is the type species of the genus of the family and infects sheep and goats, often around the mouth, resulting in acute pustular skin lesions. ORFV encodes several secreted immunomodulators including a broad-spectrum chemokine binding protein (CBP). Chemokines are a large family of secreted chemotactic proteins that activate and regulate inflammation induced leukocyte recruitment to sites of infection.

Orf virus inhibits interferon stimulated gene expression and modulates the JAK/STAT signalling pathway.

Interferons (IFNs) play a critical role as a first line of defence against viral infection. Activation of the Janus kinase/signal transducer and activation of transcription (JAK/STAT) pathway by IFNs leads to the production of IFN stimulated genes (ISGs) that block viral replication. The Parapoxvirus, Orf virus (ORFV) induces acute pustular skin lesions of sheep and goats and is transmissible to man.

Infection with recombinant orf viruses demonstrates that the viral interleukin-10 is a virulence factor.

Orf virus is the prototype parapoxvirus that causes the contagious skin disease orf. It encodes an orthologue of the cytokine interleukin (IL)-10. Recombinant orf viruses were constructed in which the viral interleukin-10 (vorfIL-10) was disabled (vorfIL-10ko) and reinserted (vorfrevIL-10) at the same locus and compared to wild-type virus for their ability to induce skin lesions in sheep.

Orf virus interleukin-10 inhibits cytokine synthesis in activated human THP-1 monocytes, but only partially impairs their proliferation.

The sheep parapoxvirus orf virus (ORFV) induces acute, pustular skin lesions in humans. ORFV encodes an orthologue of interleukin-10 (IL-10) that, whilst it closely resembles ovine IL-10 (91 % amino acid identity), shows only 75 % amino acid identity to human IL-10 (hIL-10). The anti-inflammatory potential of ORFV IL-10 in human ORFV infection was investigated by examining its immunosuppressive effects on THP-1 monocytes.