Timothy D. Smith

**W*** Wagner Rd

Warrenville, IL 60555

630-***-****

acd8jw@r.postjobfree.com

OBJECTIVE:

I am seeking a research job in the field of micro/molecular biology. I have Master's in Science in Biology

with a concentration in Biophysics and Biochem. from Northern Illinois University. During my time as a

graduate student I have learned many molecular techniques and worked with DNA, RNA, and protein

analysis. Working with numerous projects has fine-tuned my ability to multi-task and produce results. I

am organized and able to keep good documentation of all protocols followed and write up reports based

on data collected. My strong work ethic and passion for working in the scientific community I feel would

make me a great benefit to any scientific team.

RESEARCH EXPERIENCE:

Thesis Research Work: “The mtfA Transcription factor gene controls morphogenesis, gliotoxin

production and protease activity in the opportunistic human pathogen Aspergillus fumigatus.”

• Fungal Strain Development utilizing fusion PCR, Molecular Cloning, and PEG mediated

fungal transformation of Aspergillis spp. (Strains included deletion, complementation and

over-expression)

• Morphological studies include morphogenesis and asexual development of different

strains utilizing different media types and microscopy plus a hemacytometer for spore

quantification.

• Nuclear localization of MtfA by generating a strain tagged with Gfp and viewing it under

a fluorescent microscope.

• Protease analysis using an azocasein assay to determine semi quantitative protease

activity. (Looked at both total culture protease activity and secreted protease activity)

• Toxin analysis of the mycotoxin gliotoxin which involved culturing the fungus along with

extracting the toxin, followed by utilizing HPLC to compare different levels of gliotoxin

across the strains.

• Gene expression analysis looking at whether the master transcription factor played a role

in regulating different fungal global regulators, genes in the asexual development

pathway and genes within the gliotoxin gene cluster.

o Total RNA isolation from fungal cultures.

o DNAse treatment and cDNA synthesis of RNA.

o qRT-PCR analysis to find relative gene expression

Work as a Graduate Research Assistant: Protein-protein interaction in Aspergillus nidulans.

• Generate a fungal strain with the desired protein tagged with S-tag using fusion PCR and

PEG mediated transformation.

• Confirm strain using diagnostic PCR, Southern Blot Analysis, and Western blotting using

an Anti-S-tag primary antibody.

• Protein pull-down to isolate proteins that are interacting with the targeted protein.

• Prepare and send samples for proteomics.

First Author Research:

Smith TD and Calvo AM. 2014. The mtfA Transcription Factor Gene Controls

Morphogenesis, Gliotoxin Production and Virulence in the Opportunistic Human

Pathogen Aspergillus fumigatus.Euk Cell. [Epub ahead of print]

• Publication as a result of my thesis work.

• Included a virulence study I did using a Galleria mellonella(wax moth) infection

model that is widely used in the fungal and bacterial community.

Third Author Research:

Duran RM, Gregersen S, Smith TD, Bhetariya PJ, Cary JW, Harris-Coward PY, Mattison

CP, Grimm C, Calvo AM. 2014. The role of Aspergillus flavusveA in the production of

extracellular proteins during growth on starch substrates. ApplMicrobiolBiotechnol. 4

[Epub ahead of print]

• Performed the protease activity part of this paper determining the role the global

regulators veA and laeA play using an azocasein assay.

SKILLS:

• •

Aseptic Techniques Toxin Extraction

• •

Lab Safety Procedures HPLC Analysis

• •

DNA/RNA isolation Fluorescent Microscopy

• •

Fragment Purification Galleria mellonellamodel of

• infection

Primer Design

•

• Numerous Bioassays

Polymerase Chain Reaction

•

(PCR) Experimental troubleshooting

• •

Gel Electrophoresis Fungal/Bacterial

• Transformations

cDNA synthesis

•

• Grant Writing

qRT-PCR

•

• Manuscript Writing

Protein Isolation

•

• IACUC Training with Handling

Protein Pull-down

mice in a Lab Setting

• Southern/Northern/Western Blot

• Restriction Digestions

• ELISA

• Molecular Cloning

EDUCATION:

University of Illinois at Bachelors of Science Chicago, IL

Chicago Biology

• Minor in Chemistry

Northern Illinois Masters of Science (Thesis Masters) DeKalb, IL

University Biological Sciences

01/02/2012 - • Concentration on Biochem. and Biophysics

05/09/2014

ACHIEVEMENTS:

Graduate Grant Funding: I had to prepare a full research grant proposal and submit it to

•

the graduate school. From the pool of applicants only a few where selected for funding.

My grant proposal was awarded funding. (03/03/2014)

ALLFUN Scholarship: I was awarded a scholarship for ALLFUN to attend the 6th

•

Advances AgainstAspergillosis conference in Madrid, Spain. (03/01/2014)

Conference Speaker: My abstract submitted to the international 6th Advances

•

AgainstAspergillosis meeting was deemed by the Scientific Community to be of such

importance as to be selected for an oral presentation. (03/01/2014)

• Successfully wrote and defendend a Master’s Thesis entitled “The mtfA Transcription

factor gene controls morphogenesis, gliotoxin production and protease activity in the

opportunistic human pathogen Aspergillus fumigatus.” (April 2014).

Contact this candidate