The presence of a non-B-Z-structure enhances an enzyme's reactivity at its recognition site
Master of Science (MS)
First Advisor's Name
Stephen A. Winkle
First Advisor's Committee Title
Second Advisor's Name
Third Advisor's Name
Kenneth G. Furton
Enzymes, DNA, Structure
Date of Defense
Alternating (CG) sequences form an unusual conformation in the presence of cobalt hexamine. The oligomer, BZ-IV, containing a (CG)4 run (BZ-IV sequence: 5'TCGACGCGCGCGATCAGTCA- 3') was inserted at the Sal I site of the Escherichia coli pGEM-5zf(+) plasmid producing the plasmid pCW001. Hinf I digestion of pCW001 produced a 367 base pair (bp) fragment containing the BZ-IV insert. For controls, the 452 bp Hinf I fragment from the pCW001 plasmid and the 347 bp Hinf I fragment from the pGEM plasmid were used.
Digestion studies were performed using the restriction enzymes Bgl I, EcoRV, Hha I, Mbo I, Not I, Pst I, and Taq I and methylation studies were performed using dam methylase. Data were obtained by beta scanning or ethidium bromide staining the polyacrylamide gels of the digestion or methylation products. The results show that in the presence of 100 uM cobalt hexamine, in which BZ-IV takes on a non-B-Z-structure, the enzyme's ability to react and cleave its recognition site is enhanced.
Aloyo, Maria C., "The presence of a non-B-Z-structure enhances an enzyme's reactivity at its recognition site" (1992). FIU Electronic Theses and Dissertations. 1114.
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