INFORMATION TO USERS While the most advanced technology has been used to photograph and reproduce this manuscript, the quality of the reproduction is heavily dependent upon the quality of the material submitted. For example: • Manuscript pages may have indistinct print. In such cases, the best available copy has been filmed. • Manuscripts may not always be complete. In such cases, a note will indicate that it is not possible to obtain missing pages. • Copyrighted material may have been removed from the manuscript. In such cases, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, and charts) are photographed by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps. Each oversize page is also filmed as one exposure and is available, for an additional charge, as a standard 35mm slide or as a 17”x 23” black and white photographic print. Most photographs reproduce acceptably on positive microfilm or microfiche but lack the clarity on xerographic copies made from the microfilm. For an additional charge, 35mm slides of 6”x 9” black and white photographic prints are available for any photographs or illustrations that cannot be reproduced satisfactorily by xerography. Order Number 8726807 Characterization of plasmids from Bacillus thuringienais var. israelensis Clark, Burton David, Ph.D. The Ohio State University, 1987 UMI SOON.ZeebRA Ann Arbor, MI 48106 PLEASE NOTE: In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a check mark V 1. Glossy photographs or pages. y 2. Colored illustrations, paper or print ______ 3. Photographs with dark background ^ 4. I llustrations are poor copy ______ 5. Pages with black marks, not origin^ copy _ 6. Print shows through as there is text on both sides of page. 7. Indistinct, broken or small print on several pages 8. Print exceeds margin requirements _____ 9. Tightly bound copy with print lost in spine ______ 10. Computer printout pages with indistinct print. 11. Page(s) ■■ lacking when material received, and not available from school or author. 12. Page(s) seem to be missing in numbering only as text follows. 13. Two pages numbered . Text follows. 14. Curling and wrinkled pages 15. Dissertation contains pages with print at a slant, filmed as received ________ 16. O ther _________ ___________________________________________ University Microfilms International CHARACTERIZATION OF PLASMIDS FROM BACILLUS THURINGIENSIS VAR. ISRAELENSIS DISSERTATION Presented in Partial Fulfillment of the Requirements for the Doctor of Philosophy in the Graduate School of the Ohio State University By Burton David Clark, B.Sc. ***** The Ohio State University 1987 Dissertation Committee: Approved by D. H. Dean B. R. Oakley W. R. Strohl Adviser Department of Microbiology To My Wife %% ACKNOWLEDGEMENTS I would like to thank my advisor, Prof. Donald H. Dean, for his support and encouragement. Don gave me the freedom to make my own mistakes (a privilege I never hesitated to use), but was always available with advice and support when I needed it. I admire his caring for the people in his group beyond caring about their science. I would also like to thank the members of my thesis committee. Prof. Berl R. Oakley and Prof. William R. Strohl for their suggestions and advice. I am grateful to Tom Boyle, Bill Chu, Steve Cohen, Dan Ellis, John Hopper, Inara Ladzins, John Lohr, Phyllis and Bill Martin, Jim McLinden, Penny Parker, John Perkins, Fred Perlak, Laura Pettine, Josanne Sabourin, Wes Workman, Dan Zeigler, and other members of the lab whose ideas and encouragement were invaluable, and whose tolerance of my humor was admirable. I am also grateful to Philip Auron at MIT who has been patient and supportive of my completing this thesis. I want to thank my mother for her interest in my work and for instilling in me a desire to always do my best. Most of all, I want to thank my wife and best friend, Molly, for her love, devotion, and encouragement, and for dragging me away from the laboratory when I needed it most. %%% VITA February 6,1957 .................................. Born - Beverly, Massachusetts 1975-1979 ............................................. B.S., Microbiology, University of Massachusetts, Amherst, Massachusetts 1977-1979 ............................................ Undergraduate research in molecular genetics Department of Microbiology University of Massachusetts, Amherst, MA 1979-present ....................................... Graduate Research Associate, Department of Microbiology The Ohio State University, Columbus, Ohio 1985-present ........................................ Research Associate, Massachusetts Institute of Technology, Cambridge, Massachusetts. PUBLICATIONS & ABSTRACTS • Clark, B.D., Lohr, J.R., and Dean, D.H. (1980). Genetic manipulation and phenotypic properties of plasmids of the insect pathogen. Bacillus thuringiensis. Second American Society for Microbiology Conference on Genetics and Molecular Biology of Industrial Microbiology. Bloomington, ID. October 7, 1980. (abstract) * Clark, B.D., Perlak, F.J., Chu, C.-Y., and Dean, D.H. (1982). The Bacillus thuringiensis genetic systems. In: Comparative Pathobiology, edited by LA. Bulla, Jr. and T.C. Cheng, pp. 155-173. Vol. 5. Plenum Press, New York., %v • Clark, B.D., Chu, C.-Y., Cohen, S.B., and Dean, D.H. (1982). Plasmid associated toxicity in Bacillus thuringiensis var. israelensis. American Society for Microbiology Annual Meeting. Atlanta, G A. March 7, 1982. (abstract) • Dean, D.H., Clark, BJ)., Lohr, J.R., and Chu, C.-Y. (1982). Recent advances in the genetics of research in Bacillus thuringiensis. Invertebrate Pathology and Microbial Control: Proceedings of the Third International Colloquium on Invertebrate Pathology. University of Sussex, Brighton, United Kingdom, (abstract) • Clark, B.D., and Dean, D.H. (1983). A high molecular weight plasmid is associated with toxicity in Bacillus thuringiensis var. israelensis. American Society for Microbiology Annual Meeting. New Orleans, LA. March 9, 1983. (abstract) • Clark, B.D., and Dean, D.H. (1984). The antimicrobial susceptibility of various Bacillus thuringiensis var. israelensis strains. American Society of Microbiology Annual Meeting. St. Louis, MI. March 7, 1984. (abstract) • Boyle, T.M., Clark, B.D., and Dean, D.H. (1984). Restriction endonuclease mapping of three plasmids from Bacillus thuringiensis var. israelensis. Annual Meeting of the Society for Invertebrate Pathology. Davis, California. August 9,1984. (abstract) • Clark, B.D., Boyle, T.M., Chu, C.-Y., and Dean, D.H. (1985). Restriction endonuclease mapping of three plasmids from Bacillus thuringiensis var. israelensis. Gene 36, 169-171. • McLinden, J.H., Sabourin, J.R., Clark, B.D., Gensler, D.R., and Dean, D.H. (1985). Cloning and expression of an insecticidal K-73 type crystal protein from Bacillus thuringiensis var. kurstaki into Escherichia coli. Applied and Environmental Microbiology 50, 623-628. • Clark, B.D., Rosenwasser, L.J., Webb, A.C., Irie, S., Gehrke, L., and Auron, P.E. (1986). Expression of biologically active human interleukin V 1 subpeptides by transfected COS cells. Sixth International Congress of Immunology. Toronto, Canada. July 7, 1986. (abstract) • Fenton, M.J., Clark, B.D., Collins, K., Rich, A., Webb, A., and Auron, P.E. (1986). Expression of the human IL-1 beta gene in THPl cells following LPS stimulation. Sixth International . Congress of Immunology. Toronto, Canada. July 7, 1986. (abstract) • Rosenwasser, L.J., Webb, A.C., Clark, B.D., Irie, S., Chang, L., Dinarello, C.A., Gehrke, L., Wolff, S.M., Rich, A., and Auron, P.E. (1986). Expression of biologically active human interleukin 1 subpeptides by transfected simian COS cells. Proc. Natl. Acad. Sci. USA 83, 5243-5246. • Clark, B.D., Collins, K.L., Gandy, M.S., Webb, A.C., Auron, P.E. (1986). Organization and structure of the human prointerleukin 1 beta gene. Congress on Research in Lymphokines and Other Cytokines. Boston, MA. August 12, 1986. (abstract) • Fenton, M.J., Clark, B.D., Collins, K.L., Webb, A.C., and Auron, P.E. (1986). Transcriptional regulation of the human prointerleukin 1 beta gene. Congress on Research in Lymphokines and Other Cytokines. Boston, MA. August 12, 1986. (abstract) • Clark, B.D., Collins, K.L., Gandy, M.S., Webb, A.C., and Auron, P.E. (1986). Genomic sequence for human prointerleukin 1 beta: possible evolution from a reverse transcribed prointerleukin 1 alpha gene. Nucleic Acids Res. 14, 7897-7914. • Rosenwasser, L.J., Auron, P., Gehrke, L., Clark, B., McDonald, B., Bradley, B., Epstein, E., Collins, K., Webb, A. (1986) Interleukin 1 - Review of structure and function, definition of an active site for T cells, and production by cultured vascular endotheliuni. In Biologically based immunomodulators in the therapy of rheumatic diseases, Pisetsky, P.S. and Rosenwasser, L.J., eds. Elsevier Scientific Publishers, NY. m • Fenton, M.J., Clark, B.D., Collins, K.L., Webb, A.C., and Auron, P.E. (1987). Transcriptional regulation of the human prointerleukin 1 beta gene. J. Immunol. 138, 3972-3979. Fenton, M.J., Clark, B.D., Collins, K.L., Webb, A.C., and Auron, P.E. (1987). The human proIL-l;9 gene is regulated by two distinct receptor, pathways. Fifth International Lymphokine Workshop, January 1987, Clearwater, FL. (abstract) • Fenton, M.J., Clark, B.D., Alexander, S.J., Webb, A.C.j Auron, P.E. (1987). Differential expression of the human prointerleukin 1 beta gene. In: Mechanisms of Control of Gene Expression, UCLA Symposia on Molecular and Cellular Biology, Vol. 67. Collen, B., Gage, L., Siddigui, M., Skalka, A.-M., and Weissback, H., eds. Alan R. Liss, New York. • Webb, A.C., Collins, K.L., Snyder, S.E., Rosenwasser, L.J., Clark, B.D., Eddy, R.L., Shows, T.B., Auron, P.E. (1987). Molecular cloning, expression and chromosomal assignment of a potential inhibitor of human prointerleukin 1^ processing. Lymphokine Research, Vol 6, No. 1. (abstract) • Fenton, M.J., Clark, B.D., Collins, A.C., Rich, A., Webb, A.C., Auron, P.E. (1987) Two pathways of human prointerleukin 1)9 gene regulation by PMA and LPS.