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Protein Expression Core Laboratory

Services

The Protein Expression Core Laboratory aids intramural researchers in protein expression, from initial plasmid construction through expression optimization and protein purification. Core staff provides training in a number of protein expression techniques, as well as working directly with researchers to determine how best to meet project goals.

Background

Types of projects we do:

  • Cloning and subcloning Genes of Interest
    • Gateway cloning
    • Traditional cloning
    • In vitro mutagenesis
  • Expression of heterologous proteins in E. coli
    • Expression tests performed in multiple cell lines, using varied media formulations, and over a wide temperature range.
    • Expression tests may include use or rare tRNAs and chaperones
  • Baculovirus generation, propagation and protein expression checks in Insect cells
    • Titering of baculovirus stock using SF9ET cells
    • Long term baculovirus storage using TIPS protocols
  • Expression of proteins in transient transfected and generation of stable Mammalian cells
    • Trial include use of Cos-7, CHO-s and HEK293-f cells lines
    • CHO and HEK cell lines are tested in both monolayer and in suspension growth
    • Generation of Bacmam baculoviruses for scalable transient transfection in suspension mammalian cultures.
  • Protein refolding
    • 96 well Rapid dilution plate and high Hydrostatic pressure methods
  • Scale up of Protein expression systems
    • Scale up to 18 L for E. coli available
    • Scale up to 25 L for insect cell and suspension growth mammalian cell
  • Circular Dichroism measurements
    • Thermal melting curve via CD signal or Fluorescence can be done
  • Protein purification
  • Generating Lenti viral transfer vectors
  • Culturing and expansion of unique mammalian cell lines

NIEHS Shared and Core Facilities are available to NIH researchers.  Information for staff on utilizing these services can be found on the NIEHS Junction or by contacting the staff below.

Scientific Staff

Robert M. Petrovich, Ph.D.
Robert M. Petrovich, Ph.D.
Director, Protein Expression Core Facility
P.O. Box 12233
Mail Drop F1-02
Durham, N.C. 27709

Tel 919-541-4766
petrovi1@niehs.nih.gov
Tom Stanley, M.S.
Tom Stanley, M.S.
Contractor — Biologist
P.O. Box 12233
Mail Drop F1-02
Durham, N.C. 27709

Tel 919-541-7952
thomas.stanley@nih.gov
Molly Cook
Molly E. Cook
Biologist
P.O. Box 12233
Mail Drop C3-03
Durham, N.C. 27709

Tel 919-541-2120
Fax 919-541-7880
cook1@niehs.nih.gov
Lori Edwards
Lori L. Edwards
Biologist
P.O. Box 12233
Mail Drop F1-02
Durham, N.C. 27709

Tel 919-316-4689
Fax 919-316-4687
edwards7@niehs.nih.gov
Christine Malone
Christine C. Malone
Biologist
P.O. Box 12233
Mail Drop F1-02
Durham, N.C. 27709

Tel 984-287-3436
Fax 919-316-4687
malonec2@niehs.nih.gov