Lab Alumni
Christina Boob '01 - B.S.-level Scientist, Exygen Reseach, State College
Anna Bucher '01 - Medical School -
Melissa Fogg '01 - Masters Program, University of Delaware
Kevin Frederick '01 - Medical School - 
Mitchell Marzo '01 - Taking a year off before Med School
Amy Mayhew '01 - B.S.-level Scientist, Viromed Laboratories, Camden, NJ
Melissa Stokes '01 - B.S.-level Scientist, University of Virginia (Med school next year)
Julie Wagner '01 - Medical School - 
Deanne Greene '00 - Ph.D. Program at Medical University of South Carolina
Honors Project - Amplification and cloning of Staphylococcus aureus purQ into an expression vector; Analysis of FGAR amidotransferase sequences.
    Deanne did a great job in the lab. She worked through initial problems with cloning her gene by identifying a previously unrecognized restriction site in her PCR product that interfered with cloning. After designing new primers, she promptly amplified, cloned, and demonstrated expression of her gene. In addition Deanne applied her bioinformatics skills developed in upper-level coursework to an extensive analysis of FGAR amidotransferase sequences from a number of organisms. Her work contributed to two posters presented at the Council for Undergraduate Research National Conference. (She was also a wonderful babysitter!)
John Mazzullo '00 - Ph.D. Program at Penn State University
Honors Project - Synthesis of N-formylglycinamide ribonucleotide (FGAR).
    John did most of his work in Chriss McDonald's lab in the Chemistry Department. His strong chemistry background made him an excellent choice to synthesize the substrate for our enzyme. He also identified and analyzed the human FGAR amidotransferase sequence, designed PCR primers, isolated human RNA from white blood cells, and reverse transcribed it to cDNA but was unable to get the 4 kb RT-PCR product. We later learned that Postdocs elsewhere spent many months on this project. John presented his work at an American Chemical Society meeting at Bloomsburg University and won first prize in the Biochemistry section for his presentation.
Joshua Stutzman '00 - M.D. Program at Philadephia College of Osteopathic Medicine
Independent Study - Effects of 4-Androstene 3,17 Dione on Mouse Leydig Cells in Culture.
    Josh developed an interesting project. He wanted to get some experience with cell culture and did he ever! Problems with the CO2 line to the incubator led Josh through the always fun world of troubleshooting equipment. One he got the equipment operating properly, he obtained cells from mouse testes (ouch), and established well growing cell cultures but didn't have time to test the effects of androstenedione (of Mark McGuire fame).
David Wilson '00 - Optometry Program at Pennsylvania College of Optometry
Independent Study - Amplification and cloning of Staphylococcus aureus purL into an expression vector.
    Dave picked up the project where Jason left off and successfully cloned the Staphylococcus purL gene into the expression vector pTYB2. This work contributed to a poster presented at the Council for Undergraduate Research National Conference.
Jason Catanzaro '99 - Ph.D. Program at Wake Forest University
Independent Study - Amplification and cloning of Staphylococcus aureus purL into an expression vector.
    Jason made a good effort on his gene. He analyzed the sequence shortly after it was generated by Staphylococcus aureus genome project, designed the primers and successfully amplified the gene, but never got it into the vector. 
Rachel Lawton '99 - DVM Program at Cornell University
Independent Study - Amplification and cloning of Rhizobium etli purS into an expression vector.
    Rachael did a great job amplifying and cloning her gene. She obtained several clones with her gene which was fortunate because the first one she characterized was missing a few bases. Rachel also inaugurated "Country Music Night" in the lab when she and Kim were working.  She presented her work at the Pennsylvania Academy of Sciences meeting despite having to miss a lacrosse game.
Mark McCleland '99 - Ph.D. Program at University of Virginia
Honors Project - Mutagenesis of Canine Clotting Factor IX
    Mark was a tremendous asset to have in the lab. He began working in the lab during his sophomore year as a volunteer and was able to leverage that experience into an internship at the Institute for Human Gene Therapy at the University of Pennsylvania. He continued his work as an Honor's project using site-directed mutagenesis to optimize the canine clotting factor for expression in muscle tissue. Mark was invited back to U.Penn the summer after his Junior year and presented his work at the Pennsylvania Academy of Sciences meeting.
Kim Mistiszyn '99 - Optometry Program at Pennsylvania College of Optometry
Independent Study - Amplification and cloning of Rhizobium etli purQ into an expression vector.
    Kim was a lot of fun to have in the lab, and also very lucky! She amplified the purQ gene but had a few mishaps with ligation transformants and plasmid preps. The single plasmid prep to make it onto a gel (and run in the right direction) had her clone! A perfect example of the fact that you only need one! Kim was a co-author on a presentation at the Pennsylvania Academy of Sciences meeting.
Lori Schultz '99 - Research Technician
Independent Study - Amplification and cloning of Staphylococcus aureus purS into an expression vector.
    Lori made it the furthest of the first batch of students to amplify and clone FGAR Amidotransferase genes - she demonstrated expression of her purS-intein-chitin binding domain fusion protein. Lori did an excellent job and was very helpful to others in the lab. Lori presented her work at the Pennsylvania Academy of Sciences meeting and she was a contributor to a poster presented at the Council for Undergraduate Research National Conference.
Laura Singer '99 - DVM Program at Tufts University
Independent Study - Amplification and cloning of Escherichia coli purL into an expression vector.
    Laura worked very hard on the difficult project of cloning the 3.9 kb E.coli purL gene into the 7 kb expression vector pTYB2. She was able to get PCR product but never got recombinant clones. We suspect that the problem was either the large size of the PCR product or a problem with one of the primers. Laura did make the vector prep that several others in the lab used for their cloning. Despite this setback, Laura had an excellent understanding of the overall project and gave an OUTSTANDING  presentation on the background of the project at the Pennsylvania Academy of Sciences meeting. 
Matthew Georgy '98 - M.D. Program at Case Western Reserve 
Honor's Project - Construction of a Blue Fluorescent Protein Gene for Expression in Bacteria.  
    Originally, Matthew chose to examine potential interactions between the Type II  FGAR amidotransferase subunits using the yeast two-hybrid system. Difficulty obtaining the plasmids caused a last minute shift in focus to subcloning green fluorescent protein and  production a blue variant by site directed mutagenesis. When this work was initiated, the only blue version available was human codon optimized. We hope to use this construct for nodule competition studies with different Rhizobium strains. Both Matthew's insightful paper  and his Powerpoint Presentations are available online. 
Kathy Roberts '98 - Ph.D. Program at the University of Georgia 
Independent Study - Isolation of Rhizobium loti purine auxotrophs. 
    Kathy worked on trying to get the Rhizobium loti-Lotus model symbiosis up and running. She isolated a Streptomycin resistant derivative of the wild type R. loti but was unfortunate in that Streptomycin alone proved to be a poor selective agent against the donor E.coli that carried the mutagenic plasmids. Kathy also worked out some parameters for growing Lotus on agar slants and conducting nodulation assays. 
Diana Burley '97 - Research Technician 
Independent Study - Analysis of Legume Defense Responses Against Rhizobium.  
    Diana's project was to determine whether Rhizobium etli purine auxotrophs triggered cell wall modifications in bean that might prevent infection thread development. While the results of this study were inconclusive, we did learn quite a bit about using rhizobia expressing Green Fluorescent Protein and fluorescence microscopy to track infection thread development. Here is a nice example of a green fluorescent infection thread.
Jon Cook '97 - Pennsylvania State Police Academy  
Honors Project - Is purY a Component of FGAR Amidotransrerase? Sequence Analysis of Rhizobium pur genes. 
    During Jon's first semester of work, the E.coli purL gene encoding a single subunit form of the purine biosynthetic enzyme FGAR amidotransferase was cloned downstream of a Rhizobium pur gene promoter and tested for the ability to complement a Rhizobium purY mutant. The answer appears to be yes. Jon also sequenced the Rhizobium purY gene, and half of the yjii and purC genes.
Kevin Ferguson '97 - Teaching at South Williamsport H.S. , Sayre High School; Research Assistant in the department of Medical Oncology at the University of Colorado Health Sciences Center 
Honors Project - Analysis of Rhizobium etli Purine Biosynthetic Genes. Creation of Mutations in the Rhizobium etli yjii Gene.  
    Kevin was the first student to join my lab at Lycoming and did a very nice job getting non-radioactive DNA sequencing and the Tn1000 system for creating nested sets of clones up and running.  Kevin sequenced the Rhizobium purQ gene and half of yjii. In an attempt to determine the function of yjii, he cloned a kanamycin resistance cassette into yjii to create a non-polar mutation. This was then transferred onto the Rhizobium chromosome by homologous recombination. We have not yet identified any phenotypic differences between the yjii mutant and the wild type.
This page was created or last modified on 8/27/01 by Jeff Newman .
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© 1999 Jeffrey D. Newman