LESLIE E. SMITH

 

SUMMARY: Experienced research scientist with12+ years of “hands on” molecular biology skills.  My research focus has been in the fields of cancer research, radiation research, toxicology and genetics. These hands on skills and knowledge allow me to hit the ground running to bring a project to fruition within projected completion dates. I bring a creative approach to your research project through comprehensive literature searches and application of new techniques or technologies. I was most recently involved in producing recombinant proteins for plasmodium falciparum (Malaria) and HIV in a novel parasite based expression system. I have 5 years experience managing an radiation oncology research lab with concomitant experience supervising graduate students, laboratory technicians and teaching oncology interns. I am highly motivated team oriented person with a strong work ethic.

Technical Experience:

12+ years of hands-on molecular biology and toxicology experience including the following:

·         Apoptotic assays (TUNEL)

·         In vitro survival and forward mutation assays after exposure to toxic chemicals or radiation

·         Fluorescence In-Situ Hybridization (FISH) analysis of DNA damage and genomic instability

·         Dinucleotide repeat analysis for large and small scale loss of heterozygosity (LOH)

·         Design and engineering of vectors for protein expression

·         Expression  vector delivery

·         Standard PCR, Reverse Transcriptase PCR, Ligation-Mediated PCR, multiplex PCR, qPCR and PCR In-Situ

·         Isolation of proteins from protein lysates using nickel and cobalt-based resin columns

·         Site directed  mutagenesis and vector manipulation

·         Immunoprecipitation , Immunocytochemistry, and ELISA Assays

·         SDS-PAGE and immunoblotting

·         Isolation of DNA, RNA and protein from Human blood, mammalian cells in culture, bacteria, microbes and parasites

·         Western, Northern and Southern Blotting

·         Growth cultures of rodent and human cell lines as well as bacteria and other microbes

·         Broad knowledge of DNA damage and repair

·         High degree of personal responsibility and strong work ethic

·         Leadership, coaching and training skills

·         Logic, problem solving, organization and multitasking skills

·         Creative solutions to scientific problems

·         Literature searches for pertinent data and new technologies for application

·         Excellent written and oral communication

·         Extensive experience on database searching and record keeping

·         Working knowledge of sequencing and protein expression software

·         Knowledge of Microsoft Office Suite and Adobe Creative Suite

 

 

Education:

PhD in Environmental Toxicology/Molecular Biology, University of California, Riverside. 1996

Dissertation Title: Mechanisms of Genetic Instability in Heterochromatic Hotspots on Chromosome 1, 9, and 16 Exposed to 2, 6-Diaminopurine

 

Bachelor of Arts in Biology, George Washington University, Washington, DC. 1984

 

Sarah Lawrence College, Bronxville, NY. Dance Major 1981

 

Consulting Experience:

·         Served as a scientific advisor for graduate students working on individual research projects. Broad knowledge of DNA, damage, repair and apoptosis in involved in chromosomal instability and altered gene expression. Six graduate students achieved their PhD degrees under my supervision

·         Consultant for ELISA and fluorescent assays in Radiation Oncology Research and Biochemistry labs at the University of MD.

·         Scientific consultant for film “Project Metal Beast”, staring Kim Delaney in Hollywood, CA

Professional Experience:

 

Post-doctoral Research Fellow

 School of medicine, University of Maryland, 701 E. Pratt St. Baltimore, Maryland. 2010-present.

·         Recombination assays of truncated Perkisus marinus genes cloned into reporter constructs containing fluorescent markers such as green  and red tomato fluorescent proteins

·         Transfection of gene constructs into P. marinus by electroporation

·         Isolation of his-tagged proteins from parasite protein lysates using nickel and cobalt-based resin columns

·         Development of expression vectors containing highly expressed P. Marinus genes and a green fluorescent protein marker

·          Design and engineering of expression vectors for plasmodium and HIV genes

·         Western Blot analysis of expressed protein

·         Analysis of ab1 sequencing data using Sequencher 4.2 software

·         Immunocytochemistry of transfected gene constructs

 

 Research Scientist II

Opgen Inc., 708 Quince Orchard Road, Gaithersburg, MD, 2008-2009.

·         Ordered equipment and general supplies for startup research laboratory

·         Designed laboratory plan or layout for maximum efficiency and safety

·          Compared growth of measurements E. Coli K12 bacteria in Luria Broth (LB) versus Brain Heart  Infusion (BHI) media using standard Optical Density reader  and Turbidimeter using McFarland Standards

·         Optimized concentration of intermediates Standards for Optical Mapping of Bacterial genomes.

·          Investigated yield and length of genomic DNA isolated from commercial kits

·         Utilized and maintained fluorescent microscopes

·          Created an optical map of E.Coli K12.

 

Owner

 Smith Designs and It’s Art, Hon!, 3629 2nd Avenue, Edgewater, MD. (www.smithdesigns.org), 2005—2010.

·          Designed and manufactured fused glass vessels and jewelry

·         Designed and created whimsical Baltimore based Art

·         Attended 25 retail shows and 2 wholesale shows per year

 

Research Scientist and Lab Manager

University of Maryland, 655 W. Baltimore St. Bressler Research Building, Baltimore, MD. 2000 – 2005.

·         Coached, Mentored, and supervised 6 graduate students working on their Thesis projects. Managed and trained two technicians.

·         Instituted good laboratory practices (GLP) in an academic research laboratory

·         Optimized and standardized laboratory Fluorescent In-Situ (FISH) protocols

·         Maintained bacterial library of probes for all Human Chromosomes

·         Scheduled and led weekly formal Research Meetings. and Journal Club Meetings

·         Ordered laboratory equipment and supplies for a new start up lab .Designed laboratory plan or layout for maximum efficiency and safety

·         Responsible for repository of all bacteria, Human Cell lines and unstable hybrid Chinese Hamster Ovary-Human Chromosome 4 (CHOC4) cell lines

·         Utilized and maintained fluorescent microscopes

·         Maintained unstable CHOC4, HeLa , and Lymphoblastoid cells in culture

·         Judged annual graduate level Research Poster Session for the Biochemistry Department.

·         Critiqued, edited and proofread graduate student publications and peer-reviewed

  Journal articles

·         Identified mechanism of Instability in one unstable clone

·         Authored peer reviewed Review of ‘Radiation-Induced Genomic Instability:  Radiation Quality and Dose Response’ and coauthored four peer-reviewed journal articles

 

Postdoctoral Research Fellow

City of Hope, Beckman Research Institute, 1500 Duarte Rd, Duarte, CA, 1997- 2000:

·         Isolated metabolically activated forms of four polycyclic aromatic hydrocarbons (PAHs), metabolites found in cigarette smoke, using BSL3 procedures

·         Determined mutational spectra of these PAHs in the Human p53 gene using  Ligation Mediated PCR

·         Transfection of methylated and unmethylated SupF shuttle vector after exposure to 2 forms of Benzo{a}pyrene diol epoxide (BPDE)into E.Coli by electroporation or lipofection

·          Determination of Mutational spectrum of SupF vector induced by two  forms of Benzo{a}pyrene diol epoxide (BPDE)

·          Oral presentation of Mutational spectrum of Human p53 exposed to cigarette smoke metabolites at the 2000 Cancer Research Meeting. These data were solicited for publication in the Journal of the National Cancer Institute

·          Mutation data from BPDE methylated SupF vector experiment included  in a coauthored peer-reviewed journal article for Cancer Research

·          Instituted a Training Program for local High School Students

·          Conception of a SupF based project, mutational spectrum of supF following exposure to UV radiation, for a high school student’s Science Fair

·         Mentoring and training of the student during the time of this project

·         Co-authored three journal article

 

Postdoctoral Research Fellow

University of California-Riverside, Environmental Toxicology Program, 900 University Drive Riverside, CA, 1996 – 1997.

·         Optimized multiplex PCR for assessment of cancer risk in a population of pesticide workers in India

·         Trained other lab personnel in multiplex PCR and other molecular biology assays

·          Developed multiplex PCR for nucleotide repeats on Chromosome 16

·          Analyzed nucleotide repeat markers to determine the presence of uniparental disomy of Chromosome 16 in cells exposed to Diethylstilbesterol (DES)

·          Determined a method to isolate DNA for PCR amplification from metaphase spread slides stored for years at -80oC

 

Graduate Student

University of California-Riverside, Environmental Toxicology Program, 900 University Drive, Riverside California, 1990 – 1996.

·         Ordered laboratory equipment and supplies for a new start up lab

·         Designed laboratory plan or layout for maximum efficiency and safety

·         First person to clone, PCR amplify and manually sequence (Sanger) the Human APRT gene

·         Analysis of spontaneous mutations in diploid APRT gene

·         Forward Mutation analysis of hemizygous HGPRT and heterozygous TK marker loci

·          Determined Loss of Heterozygosity (LOH) induced by selection agent 2,6-Diaminopurine ((DAP) in APRT- mutants using dinucleotide repeat markers located on the q arm of  Chromosome 16

·         Confirmed extent of loss using Fluorescence in-Situ Hybridization (FISH)  for Chromosome 16 painting probes in interphase and metaphase in cells exposed to DAP

·         Utilized toxicology based micronucleus assay to determine excluded DNA material in cells exposed to DAP

·          Utilized and Maintained Fluorescent microscopes

·          Discovered mechanism of large scale  LOH using chromosome 16 painting probes,  fluorescent probes for all Human centromeres, and probes for the heterochromatin of Chromosome 16 in normal cells exposed to DAP

·          Isolated APRT+/- heterozygote for single step forward mutation analysis of APRT

·         Wrote three peer reviewed journal articles

 

Teaching Experience:

·         Radiation Biology for Oncology interns in residence. University of Maryland.  2000-2004               

·         Teaching Assistant, Molecular Biology. University of California-Riverside. 19941995.

·         Teaching Assistant, Molecular Biology. University of California-Riverside, 1993.

 

 

Accomplishments and Awards:

·         Human p53 gene data solicited for publication in the Journal of the National Cancer Institute

·         First person to clone, PCR amplify and manually sequence the human APRT gene

·         Isolation of an APRT+/- heterozygote in a human B lymphoblastoid cell line

·         Faculty nomination and service as the Student Member of the Curriculum  Affairs Committee for the University of California ETOX Program

·         Radiation Research Society Student Travel Award Recipient, 1994 and 1995.

·         Environmental Mutagen Society Travel Award Recipient, 1995

 

Publications:

Nagar S, Smith LE, Morgan WF.  Variation in apoptosis profiles in radiation-induced

genomically unstable cell lines. Radiat. Res. 2005 Mar; 163(3):324-31

 

Huang L, Grim S, Smith LE, Kim PM, Nickoloff JA, Goloubeva OG, Morgan WF. Ionizing

radiation induces delayed hyper recombination in Mammalian cells. Mol. Cell Biol. 2004 Jun; 24(11):5060-8

 

Smith LE, Nagar S, Kim GJ, Morgan WF. Radiation-Induced Genomic instability:

Radiation Quality and Dose Response. Health Phys. 2003 Jul; 85(1):23-9

 

Nagar S, Smith LE, Morgan WF. Mechanisms of cell death associated with death-inducing

factors from genomically unstable cell lines. Mutagenesis. 2003 Nov; 18(6):549-60.

 

Nagar S, Smith LE, Morgan WF. Characterization of a novel epigenetic effect

of ionizing radiation: the death-inducing effect. Cancer Res. 2003 Jan 15; 63(2):324-8

 

Yoon JH, Smith LE, Feng Z, Tang M, Lee CS, Pfeifer GP. Methylated CpG dinucleotides are the preferential targets for G-to-T transversion mutations induced by benzo[a]pyrene diol epoxide in mammalian cells: similarities with the p53 mutation spectrum in smoking-associated lung cancers. Cancer Res. 2001 Oct 1; 61(19):7110-7

 

Chen JZ, Smith L, Pfeifer GP Holmquist GP. Fluorescence-based directed termination

PCR: direct mutation characterization without sequencing. Nucleic Acids Res. 2001 Feb 15; 29(4):E17.

 

Smith LE, Denissenko MF, Bennett WP, Li H, Amin S, Tang M, Pfeifer GP. Targeting of lung cancer mutational hotspots by polycyclic aromatic hydrocarbons. J. Natl. Cancer Inst. 2000 May 17; 92(10):803-11.

 

Denissenko MF, Koudriakova TB, Smith L, O'Connor TR, Riggs AD, Pfeifer GP.

The p53 codon 249 mutational hotspot in hepatocellular carcinoma is not related to selective formation or persistence of aflatoxin B1 adducts. Oncogene. 1998 Dec 10; 17(23):3007-14

 

Smith LE, Parks KK, Hasegawa LS, Eastmond DA, Grosovsky AJ. Targeted breakage of paracentromeric heterochromatin induces chromosomal instability. Mutagenesis. 1998 Sep; 13(5):435-43

 

Pongsaensook P, Smith LE, Grosovsky AJ. Isolation of an APRT heterozygote from TK6 human lymphoblasts: predominance of multi-locus loss of heterozygosity among spontaneous APRT-mutants. Mutat. Res. 1997 Jun 9; 377(1):27-36

Smith LE, Grosovsky AJ. Genetic instability on chromosome 16 in a

human B lymphoblastoid cell line. Somat. Cell Mol. Genet. 1993 Nov; 19(6):515-27.

 

Smith LE, Grosovsky AJ. Evidence for high-frequency allele loss at the aprt

locus in TK6 human lymphoblasts. Mutat. Res. 1993 Oct; 289(2):245-54.

 

References Available upon Request