F. William Studier Biology Department, 463 Brookhaven National Laboratory Upton, NY 11973-5000 tel: (631) 344-3390 fax: (631) 344-3407 studier@bnl.gov The Group: F. William Studier (631) 344-3390 Eileen C. Matz (631) 344-3417

Research Interests:     Research has centered on conformations and interactions of DNA, molecular genetics and biochemistry of bacteriophage T7, and making T7 RNA polymerase and T7 expression signals useful for production of RNAs and proteins.     T7 has been a continuing interest since the 1960s. T7 genes were defined and mapped genetically, and functions of many of them have been determined by a variety of biochemical and physical techniques. Processes studied include entry of T7 DNA into its host cell, E. coli; overcoming host restriction; expression of T7 genes and shut-off of host functions; replication, processing and packaging of T7 DNA; and structure and assembly of phage particles. Determination of the complete nucleotide sequence of T7 DNA, 39,937 base pairs, revealed coding sequences for more than fifty T7 proteins and the arrangement of signals that direct gene expression. All of the T7 proteins can now be expressed from clones, independently or during infection, and virus mutants defective in any T7 gene have been isolated and characterized.      The challenges and outcomes of this basic research spurred the development of some widely used research methods, including sedimentation to measure size and shape of single- and double-stranded DNA, and slab gels for electrophoresis of proteins and nucleic acids. An understanding of how T7 efficiently directs an infected cell to produce T7 gene products led to the cloning of T7 RNA polymerase, a highly selective enzyme that can produce almost any RNA, and development of an expression system that uses T7 RNA polymerase and T7 expression signals to produce proteins from their cloned coding sequences in E. coli.      Recent work has concentrated on improving the convenience and reliability of the T7 expression system. Non-inducing growth media have been developed for stable maintenance of expression strains, and auto-inducing media have been developed that allow production of target proteins simply by inoculating with the expression strain and growing to saturation. These media and methods increase the convenience and reliability of protein production at any scale but are particularly useful for high throughput, because they facilitate protein production in many cultures in parallel and generally produce more target protein per volume of culture than is obtained in conventional media by induction with IPTG. An Overview & Recipes for non-inducing and auto-inducing media can be downloaded here PDF file .

Studier F.W. Slab gel electrophoresis. Trends Biochem Sci. 25: 588-590 (2000).  PubMed  Full Text Burley S.K., Almo S.C., Bonanno J.B., Capel M., Chance M.R., Gaasterland T., Lin D., Sali A., Studier F.W. and Swaminathan S. Structural genomics: beyond the human genome project. Nature Genetics 23: 151-157 (1999).  PubMed  Full Text Zhang X. and Studier F.W. Mechanism of inhibition of bacteriophage T7 RNA polymerase by T7 lysozyme. J Mol Biol. 269: 10-27 (1997).  PubMed

Cerritelli M.E. and Studier F.W. Assembly of T7 capsids from independently expressed and purified head protein and scaffolding protein. J Mol Biol. 258: 286-298 (1996).  PubMed Goldman E., Rosenberg A.H., Zubay G. and Studier F.W. Consecutive low-usage leucine codons block translation only when near the 5' end of a message in Escherichia coli. J Mol Biol. 245: 467-473 (1995).  PubMed

Cheng X., Zhang X., Pflugrath J.W. and Studier F.W. The structure of bacteriophage T7 lysozyme, a zinc amidase and an inhibitor of T7 RNA polymerase. Proc Natl Acad Sci USA 91: 4034-4038 (1994).  PubMed  Full Text PDB File:1LBA   Jmol viewer Studier F.W., Rosenberg A.H., Dunn J.J. and Dubendorff J.W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods in Enzymology 185: 60-89 (1990).  PubMed Studier F.W., and Bandyopadhyay P.K. Model for how type I restriction enzymes select cleavage sites in DNA. Proc Natl Acad Sci USA 85: 4677-4681 (1988).  PubMed  Full Text Moffatt B.A. and Studier F.W. Entry of bacteriophage T7 DNA into the cell and escape from host restriction. J Bacteriol 170: 2095-2105 (1988).  PubMed Moffatt B.A. and Studier F.W. T7 lysozyme inhibits transcription by T7 RNA polymerase. Cell 49: 221-227 (1987).  PubMed DeMassy B.,Weisberg R.A. and Studier F.W. Gene 3 endonuclease of bacteriophage T7 resolves conformationally branched structures in double-stranded DNA. J Mol Biol 193: 359-376 (1987).  PubMed Studier F.W. and Moffatt B.A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 189: 113-130 (1986).  PubMed Davanloo P., Rosenberg A.H., Dunn J.J. and Studier F.W. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci USA 81: 2035-2039 (1984).  PubMed  Full Text Studier F.W. and Dunn J.J. Organization and expression of bacteriophage T7 DNA. Cold Spring Harbor Symp Quant Biol. 47: 999-1007 (1983).  PubMed

Dunn J.J. and Studier F.W. Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol 166: 477-535 (1983).  PubMed    NCBI Sequence File of T7 DNA McDonell M.W., Simon M.N. and Studier F.W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 110: 119-146 (1977).  PubMed Studier F.W. Gene 0.3 of bacteriophage T7 acts to overcome the DNA restriction system of the host. J Mol Biol. 94: 283-295 (1975).  PubMed

Dunn J.J. and Studier F.W. T7 early RNAs are generated by site-specific cleavages. Proc Natl Acad Sci USA 70: 1559-1563 (1973).  PubMed  Full Text Studier F.W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol 79: 237-248 (1973).  PubMed

Studier F.W. Bacteriophage T7. Science 176: 367-376 (1972).  PubMed Studier F.W. Effects of the conformation of single-stranded DNA on renaturation and aggregation. J Mol Biol 41: 199-209 (1969).  PubMed Studier F.W. Sedimentation studies of the size and shape of DNA. J Mol Biol 11: 373-390 (1965).

Updated Nove. 27, 2008	Security Notice   Webteam   Site Map   Biology Home   BNL Home Page