# Protein Expression and Purification

> **NIH NIH P01** · UNIV OF NORTH CAROLINA CHAPEL HILL · 2024 · $178,500

## Abstract

SUMMARY
The Protein Expression and Purification Core will serve all Program Project investigators. Core B will express
and purify proteins for Projects 1, 2, 3, and 4. The Core Director, Dr. Sylvie Doublié, has 25+ years experience
in protein expression and purification. She will oversee the protein production aspect of the Core and directly
supervise the staff in charge of protein expression and purification. Her lab was the first to engineer a construct
of the polymerase domain of human DNA Polymerase θ that expresses well and is active. Dr. Rick Wood, the
Co-Director, has extensive experience in the expression of human proteins in eukaryotic expression systems.
His group was the first to express full-length, active human Pol θ.
Core B will produce soluble proteins and their variants for Projects 1, 2, 3, and 4. Protein constructs will be
expressed in different E. coli strains using protocols and methods, such as autoinduction, that have proven
success. The Core will also optimize the solubility and stability of proteins and complexes to be used in
biochemical and cellular assays, crystallization experiments and single-molecule studies, by characterizing
solvent effects on protein aggregation properties using dynamic light scattering and analytical gel filtration. Full
length Pol θ and other human repair factor that cannot be expressed in E. coli will be expressed in human 293
cells.
The Core is well equipped to express and purify proteins. Core B is also equipped for tissue culture work,
having the necessary incubators, hoods, microscope, centrifuge, and storage facilities.
The services provided by Core B are essential for each Project and for the success of the Program Project as
a whole. The Protein Expression and Purification Core will allow the different projects to achieve their research
goals in a cost-effective manner and with unified quality control. Core B does not duplicate any University core
facilities that would be available to our research team.

## Key facts

- **NIH application ID:** 10868630
- **Project number:** 5P01CA247773-05
- **Recipient organization:** UNIV OF NORTH CAROLINA CHAPEL HILL
- **Principal Investigator:** Sylvie Doublie
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $178,500
- **Award type:** 5
- **Project period:** 2020-07-01 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10868630

## Citation

> US National Institutes of Health, RePORTER application 10868630, Protein Expression and Purification (5P01CA247773-05). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10868630. Licensed CC0.

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