Virology
Welcome to the Molecular Virology Support Core (MVSC) website!The MVSC is a research support core at the Oregon National Primate Research Center (ONPRC), Oregon Health & Science University (OHSU) West Campus. The mission of MVSC is to provide essential virology services, reagents, specialized expertise and training to investigators in order to advance a wide array of research, with a particular focus on non-human primates (NHP). These range from basic studies in viral pathogenesis and immunology all the way to viral vaccines, gene transfer, and gene therapy.
The MVSC provides specialized expertise and infrastructure in the following specialty areas:
The MVSC has considerable experience and expertise in both viral diagnostics and production, particularly for AIDS and CMV pathogenesis and vaccine studies, and in adenoviral vector production.
- Production of virus stocks, virus vectors, and viral antigens
- Virus identification and quantification in clinical specimens
- Serologic assays to assess host antiviral immune responses
- Maintenance of cell stocks and sample processing
- Development of new reagents and standardized assays
Core Services
Adenovirus
• We ask that users clone their genetic sequences of choice into a provided shuttle plasmid
• We will then engineer and produce a custom adenoviral vector for you.
• Transgene cassette choices available:
o Inducible promoter for expression of toxic proteins
o CMV promoter and V5-tag for high-level expression and easy protein detection
o RNA interference (RNAi) cassettes for expression of short hairpin RNA (shRNA)
o Empty expression cassettes for user-supplied regulatory elements and transgenes
o Pre-made common stock vectors (such as Adeno-CMV-eGFP) also available
• Virus titration (particle titer; TCID50)
• Adenovirus replication-competent virus (RCV) assay (necessary for in vivo use)
Adeno-Associated Virus
The OHSU Vollum Viral Core will no longer be providing viral vector services at this point. We are working hard to continue offering you high quality and low cost AAV vector production services. The MVSC, in collaboration with our resident AAV expert, Dr. Hiroyuki Nakai, is developing new AAV production capabilities geared to best meet the needs of users. Although the capacity of large-scale AAV vector production is currently limited due to this transition, requests will be discussed and considered on a case-by-case basis until a new AAV vector production core is fully established. If there is in urgent need for AAV2, 7, 8, 9, DJ or other serotype vectors, please contact Dr. Christoph Kahl at kahl@ohsu.edu directly.
Lentivirus (HIV, SIV, and SHIV)
• Virus stock production
• Virus titration (sMAGI; TCID50)
• SIV plasma viral load quantification (qPCR)
• Ultrasensitive SIV detection and quantification in tissues (nested digital qPCR)
• SIV p27 protein and SIV antibody determination (ELISA)
• SIV in vitro infectivity (leukocytes)
• For lentiviral vectors, please contact disseng@ohsu.edu (Greg Dissen) at the MCB Core
Cytomegalovirus (CMV)
• Virus co-culture from animal tissues
• Virus stock and vector production
• Virus titration (TCID50)
• Crude infected cell lysate for viral protein
• Viral load quantification (qPCR)
• Antiviral antibody determination (ELISA)
• Virus plaque purification
• Virus growth curves
Expanded Simian-Pathogen Free Agents (ESPF)
• RhCMV, RRV, and SFV viral antigen preparations for ESPF screening
Vaccinia (MVA)
• Virus stock and vector production
• Virus plaque assays
Miscellaneous
• Small and large tissue processing and DNA/RNA extraction
• Virus isolation from tissues and fluids
• Manual and robotic nucleic acid isolation and purification
• Cloning and plasmid preparation
• Western blotting to confirm viral protein or transgene expression
• Cell stocks, primary and cell lines
• Sample processing and banking
The MVSC also offers data analysis and troubleshooting, custom services, consultation and training. Please inquire if you need additional assays not listed here and for assay development. We are committed to serving you and your research needs.
Contacts
Christoph A. Kahl, Ph.D., Director, kahl@ohsu.edu, 503-200-4642
Don Siess, Ph.D., Senior Research Associate, siessd@ohsu.edu, 503-629-4042
Jeffrey Torgerson, Research Assistant 2, torgersj@ohsu.edu
Fax: 949-607-2608
Shipping Address
Molecular Virology Support Core (MVSC)
Research Building, Room 046
Oregon National Primate Research Center
Oregon Health & Science University
505 NW 185th Avenue
Beaverton, OR 97006
Facilities
The MVSC laboratories are located in the Research Building centrally located on the ONPRC campus. The MVSC laboratory (over 1,000 sq. ft.) contains a tissue culture room, as well as molecular virology and general laboratory space. The laboratory is divided into a Biological Safety Level (BSL)-2 space and a separate BSL-2+ laboratory (315 sq. ft.) used for work with infectious HIV and SIV samples. The laboratories are well equipped for virus isolation/propagation, PCR-based quantitative virology methods, serology/immunology, and molecular virology under BSL-2 and BSL-2+ containment levels. Major MVSC equipment includes biosafety cabinets, fume hood, CO2 incubators, fluorescent microscope, tissue grinders, robotic nucleic acid purification system, nanodrop spectrophotometer, table-top and ultracentrifuges, PCR hood, multifunction plate reader and washer, thermocyclers for quantitative and standard PCR, bacterial incubator and shaker, liquid nitrogen dewer, fridges and freezers for sample storage.
