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Molecular Genetics Resource Laboratory

The Molecular Genetics Resource Laboratory has been set up to help investigators examine gene expression as well as to clone and express genes principally from prokaryotic organisms. The laboratory contains a variety of electrophoretic and related equipment to facilitate the extraction and characterization of DNA, RNA and proteins.

A. Instrumentation

  • Agarose gel holders and power packs
    Mini, midi, and maxi gels for the separation of DNA by electrophoresis
  • Microcentrifuge
    15,000 x G centrifuge for differential separation of cell extracts
  • Water baths
    3 variable speed and shaking for the growth of bacterial cells
  • Pulse field gel electrophoresis apparatus
    For the separation of large fragments of DNA, including chromosomes
  • Semi-automated plasmid extractor
    Permits a hands-off protocol for the isolation of up to 24 plasmids simultaneously
  • Hybridization oven
    Can hold up to 6 glass cylinders at constant temperatures for the processing of membranes using either Southern or Northern blotting procedures
  • Protein gel electrophoresis apparatus
    For separation of proteins based on mass and charge and for the processing of membranes using immunoblotting procedures
  • Auxiliary equipment
    Class II transfer hood, stationary water baths, shaking platforms, computer hooked to ethernet for access to DNA analysis software on server

B. Technical Expertise

  • Microbial cell growth including 15 liter fermentor
  • DNA extraction and purification, including both genomic and plasmid DNA.
  • Restriction mapping, subcloning, gene amplification (PCR & RT-PCR)
  • Gene expression by Northern, Western blotting and microarrays.
  • Genetically engineered vaccines using Vaccinia, Salmonella, and Brucella.
  • Protein characterization by gel electrophoresis.

C. Using the Immunotoxicity Risk Assessment Laboratory

Persons interested should first contact Dr. Stephen Boyle at 1-4641, or the Laboratory Specialist Senior, at 1-4155. Limited technical support is available for techniques used routinely; the staff and faculty will be willing to train personnel to carry out the routine recombinant DNA and protein over expression techniques. Researchers are expected to purchase all reagents and supplies necessary for carrying out techniques.

D. Projects Currently in Progress

  • Assessment of genetically engineered feline herpes virus expressing zona pellucida antigen to induce humoral and mucosal antibodies in cats.
  • Purification of recombinant rabbit zona pellucida from E. coli and yeast; assessment of the immune responses induced in cats.
  • Heterologous antigen expression in Brucella abortus RB51 to protect against anthrax and plague using a mouse model.
  • Host-pathogen interaction using microarrays to study Brucella-host interactions at the gene level using the infected macrophage and mouse models.

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