Curriculum vitae

SUMMARY OF SKILLS

  • Microbiological techniques: sterile microbiological techniques with yeast and bacteria in a level 2 biosafety lab; bright field, contrast, fluorescence, and confocal microscopy; preparation of media and solutions.
  • Genomics and proteomics methods: high throughput genomic DNA and plasmid purification; T-DNA genotyping; yeast synthetic genetic array screening and FPLC; high throughput cloning/subcloning of protein expression constructs using the Gateway cloning system; PCR amplification; Sanger sequencing with dye terminators; restriction enzyme digests; agarose gel electrophoresis; SDS- and 2D-PAGE; protein purification; western blot.
  • Bioinformatics and computer skills: facility with BLAST, MEGA, Clustal, PHYLIP, GeneMark, ExPASy, MASCOT, Dialign2, MAFFT, MEME and CLC Genomics Workbench; facility with Apple and Microsoft operating systems; Perl scripting; statistical analysis; website development; facility with creating publication-ready figures and graphics in Adobe Photoshop and Adobe Illustrator.
  • Plant biology techniques: growth and maintenance of experimental plants in controlled growth facilities; spray and syringe infiltration of Arabidopsis and other Brassicaceae with bacterial pathogens for growth assays and hypersensitive response assays; propagation of transgenic Arabidopsis lines.
  • Lab management skills: budgeting and product inventory and ordering; training undergraduate and graduate students in laboratory techniques; equipment maintenance.

EDUCATION AND AWARDS

  • 2012 Honours Bachelor of Science, Biology, University of Toronto
  • 2011 Undergraduate Genome Biology Research Award, Centre for the Analysis of Genome Evolution and Function (CAGEF) at University of Toronto
  • 2011 F. Michael Barrett Award, University of Toronto
  • 2010 WHMIS, University of Toronto
  • 2010 Laboratory Biosafety Course Level 2, University of Toronto

RELEVANT WORK EXPERIENCE

2012 – Present CAGEF Sequencing Technician
Laboratory Technician, Centre for the Analysis of Genome Evolution and Function, Department of Cell and Systems Biology, University of Toronto.

  • Developed a high-throughput method for DNA extraction of Pseudomonas aeruginosa clinical isolates in order to process over 600 clinical isolates.
  • Performed Multi Locus Sequence Typing analysis of over 1000 strains of P. aeruginosa by PCR amplification and Sanger sequencing of housekeeping genes as part of a large collaborative project.
  • Helped develop a Next-Gen Sequencing method for MLST and analyzed resulting data.
  • Adapted existing qPCR methods for Neisseria spp. to determine species identification in tonsil swabs.
  • Trained incoming graduate students in standard wet lab techniques.
  • Responsible for the hiring, training and management of undergraduate research assistants.
  • Ordered consumables and equipment for lab use and maintained lab inventory and budget.
  • Maintained stocks of bacteria, seed lines, media and solutions.

2010 – 2012 Genomic library construction and Synthetic Genetic Array
Independent research project, Department of Cell and Systems Biology, University of Toronto.

  • Expressed type 3 bacterial effectors in yeast to identify conserved host targets via a high-throughput genetic screen.
  • Cloned over 120 genes of interest into a variety of expression constructs in the Gateway cloning system and transformed these into E. coli and S. cerevisiae.
  • Used QuikChange site-directed mutagenesis to test functional dependence of putative active sites.
  • Produced and maintained a library of ~400 cloned strains in E. coli.
  • Confirmed all clones via Sanger sequencing, restriction digest and PCR.
  • Purified His- and GST-tagged proteins from E. coli constructs and assayed expression via western blot.
  • Aided in the development of a high throughput yeast plasmid transformation method and modified synthetic genetic array (SGA) screen to explore host-pathogen interactions.
  • Computationally explored protein interaction network data for relevant biochemical pathways identified in the SGA screen.

May – Aug 2010 Hypersensitive Response in Brassicaceae
Volunteer research project, Department of Cell and Systems Biology, University of Toronto.
Brassicaceae response to injection by different pathogenic strains of Pseudomonas syringae.

  • Maintained plant stocks according to controlled experimental protocols.
  • Pressure-infiltrated plant tissues with bacterial strains at various concentrations in order to evaluate differential growth patterns.
  • Evaluated plant immune response both qualitatively through hypersensitive response assays and quantitatively through post-infiltration bacterial growth assays.

May – Aug 2009 Viral control of Gypsy Moth larvae
Volunteer research project, Department of Forestry, University of Toronto.

  • Maintained clonal tree stocks in a controlled growth environment.
  • Maintained larval stocks on appropriate media.
  • Distributed larvae and assessed larval and tree health daily both pre-and post-application of virus.

PUBLICATIONS AND POSTERS

Amy AH, Middleton MA, Youn JY, Sharifpoor S, Andrews B, Desveaux D, and Guttman DS. 2014.
Identifying Conserved Host Targets of Pseudomonas syringae Type III Secreted Effectors. Poster presented at XVI International Congress on Molecular Plant-Microbe Interactions, Rhodes, Greece.

Lee AH, Middleton MA, Guttman DS, Desveaux D. 2013.
Phytopathogen type III effectors as probes of biological systems. Microbial Biotechnology 1062:301-15.

Middleton MA, Yau Y, Stanojevic S, Ratjen F, Tullis E, Speert D, Zlosnik J, Guttman DS, Waters, V. 2013.
National survey of clonal Pseudomonas aeruginosa in Canadian cystic fibrosis patients. Poster presented at the 27th Annual North American Cystic Fibrosis Conference, Salt Lake City, Utah, USA.

Lee AH, Middleton MA, Youn JY, Sharifpoor S, Andrews B, Guttman DS, and Desveaux D. 2012.
Pathogenic genetic array of bacterial effectors to screen for conserved eukaryotic pathways. Poster presented at the 30th New Phytologist Symposium: Immunomodulation by plant-associated organisms, Fallen Leaf Lake, California, USA.

Lee AH, Middleton MA, Youn JY, Sharifpoor S, Andrews B, Guttman DS, and Desveaux D. 2012.
Pathogenic genetic array of bacterial effectors to screen for conserved eukaryotic pathways. Poster presented at the 13th International Conference on Systems Biology, Toronto, Ontario, Canada.

Lee AH, Middleton MA, Youn JY, Andrews B, Guttman DS, and Desveaux D. 2011.
Identifying the conserved host targets of Pseudomonas syringae. Poster presented at the Gordon Research Conference in Cellular Systems Biology, Davidson College, North Carolina, USA.

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