The specific objectives for the current period of investigation are: I. Cloning of the BamHI R/M enzymes a. Polynucleotide synthesis and purification. b. Solution of DNA probes for cloning of BamHI restriction modification enzyme genes. c. Options for cloning of R/M genes. d. Determination of primary sequence of BamHI R/M enzymes as inferred from DNA sequences. II. Physical and biochemical studies on BamHI R/M enzymes. a. Crystallization of BamHI endonuclease b. Co-crystallization of BamHI and the cognate oligonucleotides c. Preparation of BamHI palindrome with modified flanking sequences d. Analysis of BamHI Kinetic Data e. Reaction rates of BamHI endonuclease with variations in the location of the recognition sequence. III. Studies on HinGUII R/M enzymes a. Purification of restriction modification enzymes. b. Characterization of modification enzymes(s). c. Identification of the modification acceptor.
| Hensley, P; Nardone, G; Chirikjian, J G et al. (1990) The time-resolved kinetics of superhelical DNA cleavage by BamHI restriction endonuclease. J Biol Chem 265:15300-7 |
| Connaughton, J F; Vanek, P G; Lee-Lin, S Q et al. (1988) Cloning of the BamHI methyl transferase gene from Bacillus amyloliquefaciens. Gene Anal Tech 5:116-24 |
| Nardone, G; George, J; Chirikjian, J G (1986) Differences in the kinetic properties of BamHI endonuclease and methylase with linear DNA substrates. J Biol Chem 261:12128-33 |
| George, J; Nardone, G; Chirikjian, J G (1985) Sequence-specific BamHI endonuclease. The proposed role of arginine residues in substrate binding and recognition. J Biol Chem 260:14387-92 |
