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ISOLATION AND CHARACTERIZATION OF THE MODIFIED BASE FOUND FOR BACTERIOPHAGE SN1 DNA
Abstract
Bacteriophage SN1 contains a modified base that replaced 35% of the guanine. Mass spectral studies using field desorption and chemical ionization established a molecular weight of 579. ('1)H-Nuclear magnetic resonance, infrared the ultraviolet absorption data indicate that the structure of the modified base is comprised of three chief structural components: (1) a purine ring system that is linked to, (2) aromatic or heterocyclic ring(s) via (3) an aliphatic linkage. ('13)C-nmr indicated the presence of sulfur which is supported by infrared absorption and a large M+2 peak obtained by field desorption. The presence of a five-membered ring system is strongly suggested by IR absorption. Atomic absorption analysis revealed the presence of zinc in association with the modified base and with phage SN1 DNA. Ultraviolet absorption data and agarose gel electrophoresis of bacteriophage SN1 DNA indicate a conformational change of the DNA mediated by the presence of zinc. Electron microscopy of phage SN1 DNA and resistance to S(,1) endonuclease indicate no single-stranded regions in SN1 DNA. The molecular weight of phage SN1 DNA was determined to be 42 x 10('6) daltons on the basis of agarose gel electrophoresis. Alkaline denaturation of bacteriophage SN1 DNA failed to yield the expected complementary single strands. Bacteriophage SN1 DNA was shown to be highly resistant to most Type II restriction endonucleases. No host Type II restriction system was demonstrated. The requirement of trace metals for successful phage development was shown by phage burst data. {('14)C}-amino acid incorporation and SDS-PAGE of proteins of phage infected cells also support the requirement for trace metals. Phage SN1 was shown to contain 14 structural proteins by SDS-PAGE.
Subject Area
Microbiology|Molecular biology
Recommended Citation
RICE, WILLIAM CLINTON, "ISOLATION AND CHARACTERIZATION OF THE MODIFIED BASE FOUND FOR BACTERIOPHAGE SN1 DNA" (1981). ETD collection for University of Nebraska-Lincoln. AAI8208374.
https://digitalcommons.unl.edu/dissertations/AAI8208374