Joseph Steimel

Relevant Course Work: organic chemistry, qualitative chemistry, parasitology, genetics, microbiology, vertebrate biology, ecology, botany, animal behavior, Iowa natural history and introduction to plant molecular biology (ISU).

Work Experience

Research Associate II, Plant Pathology Department, Iowa State University, Ames: 1992 to present. Teach molecular biology techniques to graduate students and visiting scientist. Supervision of hourly workers. Assist in the genetic analysis of fungal cultures and maintain culture collection. Perform: DNA isolation, PCR, DNA sequencing, RFLP'S, microsatellite analysis, genewalking, In gel hybridization, and isozymes. Purchase all supplies and equipment. Assist in the publication of research in scientific journals. Responsible for maintenance of all equipment.

Laboratory Technician II, Iowa State Seed Laboratory, Ames: September 1989 to March 1992. Establishment of a varietal identification laboratory which researched the use of seed isozymes and storage proteins as a means of maintaining quality seed lines. Interpreted and analyzed isozyme data. Performed background literature searches on varietal identification technique. Responsible for the development of laboratory protocols used in the identification of oat and wheat varieties. Responsible for the Purchase of lab equipment and supplies.

Laboratory Technician II, Horticulture Department, Iowa State University, Ames: February 1988 to September 1989. Assisted in a plant molecular biology laboratory which researched the expression of major tuber protein genes and characterized the expression of the genes as it is associated with the process of tuberization. Skills used included the isolation of DNA and RNA, western blotting, SDS PAGE and northern/southern blots. Responsible for the Purchase of lab equipment and supplies.

Polymerase Chain Reaction: Experienced in the amplification of fungal ribosomal RNA (rRNA) genes used in population genetics. These regions include the Internal transcribed spacer region (ITS), Intergenic spacer region (IGS), Nuclear large and small subunit, Mitochondrial large and small subunits. Furthermore, I am skilled in the amplification of the ceratoplatnin locus, scalactylone dehydrogenase, betatublin and mating type genes using degenerate primers. See example #1, #2.

Microsatellite Identification: Proficient in isolation techniques (anchored PCR, nylon memebrane hybridization, magnetic beads) for the identification of microsatellite loci. Experienced in the scoring of fluorescently labeled (HEX, TET, FAM) microsatellite alleles using ABI Genescan™ / Genotyper™ software. See example below.

Genewalking: Skilled in the use of both Tail PCR and ClonTechs genewalking kit for the acquisition of unknown DNA flanking regions.

DNA Sequencing: Experienced in the direct sequencing of PCR products. This includes purification of template, pouring and running of sequencing gels, The analysis of sequence data using programs such as Sequence Navigator™ and Auto Assembler™.

Cloning: Skilled in the gel purification of PCR products and the cloning of these products into T/A cloning vectors. Transformation, plasmid isolation and clone screening are additional skills.

In Gel Hybridization: Experienced in the labeling and hybridization of P₃₂labeled oligonucleotides to total genomic DNA while still in the gel matrix. See example below.

DNA Skills: Insert isolation, slot blots, restriction digest, southern and northern blotting, DNA Isolation from plant and fungal tissue, random primed labeling, and end labeling of oligonucleotides.

Isozymes: Skilled in the use of isozymes for the varietial identification of maize and soybean lines. Including the scoring of alleles and determination of parentage. Skilled in all aspects of running, pouring and staining of gels. See example below.

Polyacrylamide Electrophoresis: Experienced in the separation of proteins using both SDS PAGE and ACID PAGE. Also skilled in western blotting.

Additional Laboratory Skills: Familiar with the purchase of supplies and equipment. Skilled in the supervision and training of lab personnel. I have received safety training in chemical waste management, use of radioactive materials, and proper use of a respirator .

Computer Skills: Competent in the use of either Windows or Macintosh based operating systems. Here are a few of the common programs I am familiar with: Microsoft Word, Microsoft Excel, Microsoft Frontpage, Macromedia Freehand, Adobe Photoshop, ABI Sequence Navigator, ABI Genescan, ABI Genotyper, ABI Autoassembler, Oligo and NIH Image.

Bio-Rad Gel Documentation system: Familiar with digital imaging and the use of Bio-Rad software for molecular weight determination.

Internet Skills: Experienced in the searching and downloading of sequence data from the genebank database (BLAST). Also proficient in the submission of sequence data to genebank (Sequin).

	Restriction patterns of total genomic DNA of Heterobasidon annosum with HAE III. This Image demonstrates the use of restriction enzymes in the analysis of the mitochondrial genome. Click the image to see a 107K GIF and a more detailed explanation.
	Restriction patterns of total genomic DNA of Heterobasidon annosum with PST I. This is an example of a restriction that would be used for in gel hybridization with an oligonucleotide. Click the image to see a 107K GIF and a more detailed explanation.
	Restriction patterns of total genomic DNA of Chlara australis and several Ceratacystis species restricted with HAE III. Click the image to see a 146K GIF and a more detailed explanation.
	Isozyme patterns of several inbred lines of maize, for the enzyme malate dehydrogenase (MDH) . Click the image to see a 17K GIF and a more detailed explanation
	PCR restriction patterns of the internal transcribed spacer region (ITS) of several fungi isolated from soybean stems. Click the image to see two images demonstrating the variation observed at the species level.
	DNA sequencing of PCR amplified products. Click the image to see an electropherogram and a brief explanation.
	DNA fingerinting bands of the aggressive and mild genotypes of Phialophora gregata based on PstI digests and hybridization with ³²P-labeled oligonucleotide (CAT)₅. Approximate band sizes (kb) are indicated for the polymorphic bands. Click the image to see a larger gel image and a map showing the distribution of aggressive and mild isolates in the North Central Region growing region.
	Microsatellite alleles analyzed using the ABI Genotyper program. Click the image to see a larger image and a brief explanation of the project. These images represent ABI Genescan files in which the forward primer was fluorescently labeled (HEX,TET,FAM) and then the PCR product electrophoresised on an ABI 3700 gel system. The system at ISU is capable of detecting a single base pair molecular weight difference. The examples presented here represent trinucleotide repeats. Many of these repeats were isolated using the technique of Edwards, or by "anchored PCR". Both techniques required genewalking back across the repeat region to obtain enough of the flanking regions to design amplification primers.
	Electropherogram of a recovered microsatellite sequence. Click the image to see a larger image of the electropherogram and a brief explanation of the technique used to isolate this unique sequence.
	Protoplast of Ceratocystis fimbriata. Click the image to see a larger image of the microscope field and a brief explanation of the technique used to produce these conidia without cell walls (protoplast).

Ceratocystis Publications

C. J. B. Engelbrecht, T.C. Harrington, J Steimel, and P. Capretti. 2004. Genetic variation in eastern North American and putatively introduced populations of Ceratocystis fimbriata f. platani . Molecular Ecology 13:2995-3005. (C.F. Italy.pdf)

J. Steimel, C. J. B. Engelbrecht and T.C. Harrington. 2004. Development and characterization of microsatellite markers for the fungus Ceratocystsis fimbriata. Molecular Ecology Notes 4:215-218 (Microsat Cerat.pdf)

Harrington, T. C., N. V. Pashenova, D. L. McNew, J. Steimel, and M. Y. Konstantinov. 2002. Species delimitation and host specialization of Ceratocystis laricicola and C. polonica to larch and spruce. Plant Dis. 86:418-422. (laricicola.pdf)

Roux, J., T. C. Harrington, J. P. Steimel, and M. J. Wingfield. 2001. Genetic variation in the wilt pathogen Ceratocystis albofundus. Mycoscience 42: 327-332.

Witthuhn, R. C., T. C. Harrington, B. D. Wingfield, J. Steimel, and M. J. Wingfield. 2000. Deletion of the MAT-2 mating type gene during uni-directional mating type switching in Ceratocystis. Current Genetics 38:48-52 . (Mat deletion.pdf).

Witthuhn, R.C., T. C. Harrington, J. P. Steimel, B. D. Wingfield, and M. J. Wingfield. 2000. Comparison of isozymes, rDNA spacer regions, and MAT-2 DNA sequences as phylogenetic characters in the analysis of the Ceratocystis coerulescens complex. Mycologia 92:447-452.

Harrington, T. C., J. Steimel, and G. A. Kile. 1998. Genetic variation in three Ceratocystis species with outcrossing, selfing and asexual reproductive strategies. European Journal of Forest Pathology 28:217-218.

Harrington, T. C., J. Steimel, M. J. Wingfield, and G. A. Kile. 1996. Isozyme variation and species delimitation in the Ceratocystis coerulescens complex. Mycologia 88:104-113.

Harrington, T. C., D. M. Rizzo, R. A. DeScenzo, and J. Steimel. 1998. Genetic relationships among clones of Heterobasidion annosum in a pine forest. To be published in: C. Delatour, ed., Proc. 9th Intern. Conf. on Root and Butt Rots. INRA.

Thomas C. Harrington, Doug McNew, and Joseph Steimel. Phylogeny and taxonomy of the Ophiostoma piceae complex and the Dutch elm disease fungi. Mycologia: Vol. 93, No. 1, pp. 111-136

Leptographium Publications

Diana L. Six, Thomas C. Harrington, Joseph Steimel, Douglas Mcnew, T.D. Paine. 2003. Genetic relationships among Leptographium terebrantis and the mycangial fungi of three western Dendroctonus bark beetles. Mycologia 95 (5): 781-792. (Leptoterebr.pdf)

Armillaria Publications

James J. Worrall, Kelley F. Sullivan, Thomas C. Harrington, Joseph P. Steimel. 2004. Incidence, host relations and population structure of Armillaria ostoyae in Colorado campgrounds. Forest ecology and management 192: 191-206. (A.Colorado.pdf)

Coetzee, M. P. A., B. D. Wingfield, T. C. Harrington, J. Steimel, T. A. Coutinho, and M .J. Wingfield. 2001. The root rot fungus Armillaria mellea introduced into South Africa by early Dutch settlers. Molecular Ecology 10:387-396. (Dutch Armillaria.pdf)

Phialophora gregata Publications

T. C. Harrington, J. Steimel, F. Workneh, and X. B. Yang. 2003 Characterization and distribution of two races of Phialophora gregata in the North Central United States. Phytopathology (Gregata.pdf)

Harrington, T. C., J. Steimel, F. Workneh, and X. B. Yang. 2000. Molecular identification of fungi associated with vascular discoloration of soybean in the North Central United States. Plant Dis. 84:83-89.(SoyFungi.pdf)

Wingfield, B. D., T. C. Harrington, and J. Steimel. 1996. A simple method for detection of mitochondrial DNA polymorphisms. Fungal Genetics Newsletter 43:56-60. (Mitochondrial DNA.html).

Joseph P. Steimel

Education

Work Experience

Ceratocystis Publications

Leptographium Publications

Armillaria Publications

Phialophora gregata Publications