The Old Well
University of North Carolina,
Chapel Hill

• Education
• Certifications
• Professional Experience
• Publications
• Abstracts
• Computer Skills
• Lab Skills

Ph.D. Microbiology and Immunology, 1996
University of North Carolina,
Chapel Hill, NC

B.A. Biology, cum laude, 1987
College of the Holy Cross,
Worcester, MA

A.A.S. Database Management, High Honors, 2007
Wake Technical Community College,
Raleigh, NC

Certificate in Bioinformatics, 2003
Wake Technical Community College,
Raleigh, NC

Certificate in Computer Programming: Oracle Developer, 2004
Wake Technical Community College,
Raleigh, NC

Certificate in Computer Programming: Oracle DBA, 2006
Wake Technical Community College,
Raleigh, NC

Oracle9i PL/SQL Developer Certified Associate (OCA)

Research Associate, 2003 - 2006
Postdoctoral Research Associate, 1996 - 2003
Advisor:  Dr. Clyde A. Hutchison III
Department of Microbiology and Immunology
University of North Carolina, Chapel Hill, NC
 

• Project leader for site-directed mutagenesis of recombinant HIV-1 reverse transcriptase used to define and characterize amino acid residues involved in protein stability and subunit dimerization of the HIV-1 reverse transcriptase.  We now have a collection of over 600 single amino acid substitution mutants in the HIV-1 reverse transcriptase (click here for a current list of our mutants).  One goal of this work is to provide new insights into designing novel therapeutic agents to inhibit HIV-1 replication.

• Designed and maintained an Oracle database to store the genotype and phenotype data for 600 HIV-1 reverse transcriptase single amino acid substitution mutants generated in the Hutchison Lab. This database also contains data from analyses of HIV-1 sequences and protein structures.

• Developed an interactive web application using Perl/CGI to allow researchers to query an Oracle database containing the Hutchison's lab reverse transcriptase mutant data.

• Constructed and analyzed structural models of reverse transcriptases related to HIV-1 to study the evolution of residues involved in protein stability and folding.  Click here for the list of retroviruses used in this study.

• Evalutated amino-acid substitutions in the reverse transcriptase of clinical HIV-1 isolates using phenotype data from a collection of HIV-1 reverse transcriptase mutants.  This analysis can serve as a method to predict if reported sequences of HIV-1 isolates are biologically functional.  The mutational data can also be used to aid in the understanding of drug resistant HIV-1 reverse transcriptase mutants.

Graduate Student, 1988-96
Dissertation Advisor:  Dr. Clyde A. Hutchison III
Department of Microbiology and Immunology
University of North Carolina, Chapel Hill, NC
 

• High density saturation mutagenesis project of recombinant HIV-1 reverse transcriptase expressed in E. coli.  The phenotype data generated from a collection of 366 different single amino acid substitutions in the HIV-1 reverse transcriptase provided a "genetic" means to identify amino acid residuess that are important for protein function or protein stability.

• Phenotype data from the mutant collection were combined with crystallographic data of several HIV-1 reverse transcriptase structures as well as the structure of the evolutionarily related Moloney murine leukemia virus reverse transcriptase to identify hydrogen bonding and hydrophobic interactions critical for the protein stability of this enzyme.

• Besides providing a better understanding of the HIV-1 reverse transcriptase in particular, this project demonstrated the utility of saturation mutagenesis to study the function and stability of proteins in general, especially for proteins for which no structural data are available.

Crystal Structure of the 66 kD subunit of the HIV-1 Reverse Transcriptase

Teaching Assistant, 1989-90
University of North Carolina, Chapel Hill, NC

Taught two semesters of Microbiology 55 Lab
(General Microbiology for nursing students)

Graduate Student, 1987-88
Department of Microbiology and Immunology
University of North Carolina, Chapel Hill, NC

Four month rotation in Dr. Jack Griffith's lab where I used electron microscopy to visualize the binding of the tryptophan repressor protein (TrpR) to DNA.

Undergraduate Research Fellow, 1985-86
University of Massachusetts Medical School, Worcester, MA

Research during the summers of 1985 and 1986 in Dr. Carel Mulder's lab where I studied the molecular biology of herpesvirus saimiri, an oncogenic member of the Herpesvirus family.

Undergraduate Research Student, 1985-86
College of the Holy Cross, Worcester, MA

Three semesters of undergraduate research in Dr. George R. Hoffmann's lab where I studied the ability of various chemicals to induce genetic duplications in Salmonella typhimurium.

Wrobel, J. A., Conrad, M. J., Bloedon, E., Swanstrom, R., and Hutchison, C. A., III.  (2000).  Analysis of HIV type 1 Reverse Transcriptase:  Comparing Sequences of Viral Isolates with Mutational Data.  AIDS Research and Human Retroviruses 16, 2049-2054.  [PubMed][Supplemental material]

Wrobel, J. A., Chao, S. -F, Conrad, M. J., Merker, J. D., Swanstrom, R., Pielak, G. J., and Hutchison, C. A., III.  (1998).  A Genetic Approach for Identifying Critical Residues in the Fingers and Palm Subdomains of HIV-1 Reverse Transcriptase. Proc. Natl. Acad Sci. USA 95, 638-645. [PubMed]

Wrobel, J. A.  (1996). Saturation Mutation and Crystal Structure Analysis of the HIV-1 Reverse Transcriptase Fingers and Palm Subdomains.  Ph.D. dissertation, University of North Carolina, Chapel Hill.

Hoffmann, G. R., Sprague, K. M., Wrobel, J. A., and Wroblewski, D. H.  (1987).  A Recombinagenic Effect of Strychnine in Salmonella typhimurium.  Environmental and Molecular Mutagenesis 10, 27-33. [PubMed]

Wrobel et al.  Mutational Analysis of Residues Critical for the Protein Stability of the HIV-1 Reverse Transcriptase.  (Manuscript in preparation).

Wrobel, J. A., Bloedon, E., Conrad, M. J., Hutchison, C. A., III  (1999).  A Mutational Study of Amino Acid Residues Involved in Protein Stability and Dimerization of the HIV-1 Reverse Transcriptase.  (Abstractfor the 1999 meeting on Retroviruses, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY May 1999).

Chao, S. -F., Wrobel, J., Conrad, M., Kaplan, A. H., Swanstrom, R., Hutchison, C. A.  (1995).  Saturation Mutagenesis of a 109 Amino Acid Segment of the Reverse Transcriptase of Human Immunodeficiency Virus Type 1.  (Abstract for the 1995 meeting on Retroviruses, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, May 1995).

Hoffmann, G. R., Sprague, K. M., Van Haight, C. C., Wrobel, J. A., and Wroblewski, D. H.  (1987).  Induction of Genetic Duplications in Bacteria by b-Propiolactone and Strychnine.  (Abstract for Eighteenth Annual Meeting of the Environmental Mutagen Society, San Francisco, CA, April 1987).

Hoffmann, G. R., Sprague, K., Wrobel, J.  (1986).  Studies on a Recombinagenic Effect of Ultraviolet Light and Chemicals in Bacteria.  (Abstract for the meeting of the Mutagenesis Association of New England, Amherst, MA, May 16, 1986).

Relational Database Management Systems:  Oracle, MySQL, Microsoft Access

Programming Languages: Java (J2SE 5.0, Servlets, JSP, JDBC), Visual Basic, Perl/CGI, PL/SQL, C++, ASP.NET, VBA, JavaScript, HTML, XML

Developer Tools: Visual Studio, Eclipse, NetBeans, XML Spy, Toad for MySQL, Oracle SQL Developer, Crystal Reports

Operating Systems:  UNIX, Linux, Mac OS, Windows XP

Data Analysis Programs:  Microsoft Excel, Cricket Graph

Website Editing Programs:  Dreamweaver, Flash

Word Processing Programs:  Microsoft Word

Presentation/Drawing Programs:  Microsoft PowerPoint, MacDraw Pro

Image Editing:  Adobe Photoshop, Adobe Illustrator

Protein Crystal Structure Analysis Programs:  Kinemage, RasMol, Protein Explorer, SwissPdbViewer, SYBYL, INSIGHT II, RIBBONS, MolScript, PyMOL

Protein/DNA Analysis Programs:  GCG package, Vector NTI

Computer Education I received at the University of North Carolina:

Course #	Course Name	Semester	Year
COMP 14	Introduction to Programming (C++)	Summer	1999
BIOC 156	Biomolecular Informatics	Spring	2000
COMP 14	Introduction to Programming (Java)	Summer	2000
COMP 114	Systematic Programming	Fall	2000
COMP 121	Data Structures	Spring	2001
COMP 117	Web Programming	Fall	2001
COMP 130	Files and Databases	Spring	2002
INLS 256	Database I	Spring	2002
INLS 258	Database II	Fall	2002

• ATN Computer Training Workshops

• UNC Center for Bioinformatics Workshops

• UNC Center for Structural Biology Workshops

Computer Education I received at Wake Technical Community College (Raleigh, NC):

List of Continuing Education Courses I attended at Wake Tech

During my 21 year research career, I have acquired experience in the following molecular biology techniques:

Site-directed mutagenesis, PCR, cell culture, DNA/RNA isolation, DNA sequencing, DNA cloning, recombinant protein expression in E. coli, immunologic techniques (such as western blot analysis), bacterial assays to screen mutagens, use of the electron microscope to visualize protein-DNA interactions, computer analysis of sequence data and crystal structures of proteins.

• Hutchison Lab Reverse Transcriptase Web Page (maintained by John Wrobel)

• View my interactive web pages   

• Photos of the 1999 Retrovirus meeting at Cold Spring Harbor Laboratory

• Photos of the University of North Carolina at Chapel Hill

Page last updated on August 30, 2008