Dr Euan W. Baxter
- Position: Research Fellow
- Areas of expertise: protein interactions; boluminescence; fusion protein expression; viral vectors for stable cell lines; flow cytometry; cellular assay development
- Email: E.W.Baxter@leeds.ac.uk
- Phone: +44(0)113 343 8439
- Location: 7.01 LIGHT Building
- Website: LinkedIn | Researchgate | ORCID
Profile
My PhD at the Medical School at Edinburgh University allowed me to follow a career in Research. I was a post-doctoral Research Fellow at the Veterinary School in Glasgow for eight years, working on the development of thymic lymphoma using classical molecular techniques. We studied the genes upregulated by nearby proviral integrations in MoMLV-induced thymic lymphomas by Southern blotting and I attempted to clone new proviral integration sites from genomic libraries I made. The proviral integration sites were often near oncogenes which encoded transcription factors so I developed an interest in gene regulation.
My interests took me to the Fred Hutchinson Cancer Research Center in Seattle where I worked for almost four years. I worked on how chromatin structure is involved in gene regulation of an important locus which encodes serine protease inhibitors.
I joined the University of Leeds as a post-doctoral fellow in 2004 and worked on chromatin and gene regulation until the end of the contract in 2007. I worked in the YCR p53 Research Laboratory in the University of York in 2008 and 2009, then joined Leeds again as a post-doc in the Institute for Cancer and Pathology. I started my present position in 2013 and I have been working on the biology of Fc gamma receptors using molecular cloning and recombinant bioluminescent protein expression in cell lines.
Research interests
My interest in gene regulation took me to the Fred Hutchinson Cancer Research Center in Seattle, where I developed Chromatin Immunoprecipitation assays and applied this technique to the regulation of a cluster of Serine protease inhibitors (SERPINS) at 14q23.1. I continued to work on chromatin at Leeds, using DNAseI hypersensitive mapping to identify regulatory regions in the IL-3 / GM-CSF locus. This project involved long-term culture of mast cells, and isolation of memory T cells and thymocytes for chromatin structure studies.
Whilst working in the YCR p53 Research Laboratory at the University of York I gained experience in using siRNA to knock down gene expression. I applied this skill to the SIRT1 gene which acetylates the tumour suppressor p53 and the phosphatase WIP1 which also acts on p53 to inactivate the molecule. This work showed "oncogene addiction" in medulloblastoma cells: reducing the post translational modifications on p53 increased apoptosis in these medulloblastoma cells.I carried out apoptosis assays using Annexin V to show up cells in early-stage apoptosis. I carried out similar experiments on bladder cancer cell lines in Leeds.
I applied a range of molecular biology techniques in cloning Fc gamma receptors (FcgRs), including site-directed mutagenesis, homologous recombination cloning and transfection. I have cloned tagged versions of Fc gamma receptors and made stable cell lines expressing them in HEK293 cells. I used these to show the specificity of protein Affimers in assays I developed using FACs. These reagents showed very fine specificity and could displace the normal ligand from the receptor. I also have some experience in the differentiation of monocytic THP-1 cells into distinct macrophage subtypes and assessment of marker transcripts. I have just made some vectors to encode tagged versions of FcgRs, and the adaptor molecules which bind to the Immuno tyrosine motifs (ITAMs) on the intracellular side of the FcgRIIa. These fusion proteins form a complete luciferase enzyme when they are in close apposition and so show when signalling takes place in the transfected cells. I have been studying the kinetics of this process in different receptor idiotypes.
Qualifications
- PhD
- BSc (Hons)
Research groups and institutes
- Discovery and Translational Science