A restriction map of DNA from bacteriophage P3-C1 for the endonucleases BamH I, Bgl II, EcoR I, Hind III, Sac I and Sca I is described in this study. The phage DNA was determined to be a linear double stranded molecule with a unit genomic length of 31 kb. After electrophoresis in 0.5% agarose gels, the undigested DNA appeared as three molecules of 31, 62 and 93 kb in length. The Hind III digestion pattern of the individual 31 kb molecule was identical to the 62 kb and 93 kb Hind III pattern. With all enzymes except Hind III, two fainter bands were consistently present. A band of normal intensity was present in each digest with the size that was equal to the sum of the lengths of the faint fragments of the same digestion pattern. In the double digestion patterns, there were extra bands that occupied the same region in which the faint bands had been mapped.
In order to account for all the observations, the restriction map was based on the two larger molecules of the native DNA. All fragments of the single and double digestion patterns could be explained by this map. The faint bands always occupied the terminal portions of the two unit length concatemeric molecule and the fragment equal to the sum of the two faint bands was found to occupy the homologous region at the junction of the two genomes constituting the concatemeric molecule. It is possible that the faintness of the fragments mapped to the terminal regions is due to their lower molar yield as a consequence of P3 DNA being in the concatemeric forms.
It was found that the faint EcoR I C fragment could hybridize to all the fragments from the six digestion products. However, the normal Hind III C fragment hybridized only to bands of the same DNA region. These observations seem to be unique to P3.
Leonard R. Bullas
George T. Javor
Barry L. Taylor
Anthony J. Zuccarelli
Master of Science (MS)
Year Degree Awarded
Date (Title Page)
Library of Congress/MESH Subject Headings
Bacteriophages -- analysis; DNA Viruses -- analysis
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This title appears here courtesy of the author, who has granted Loma Linda University a limited, non-exclusive right to make this publication available to the public. The author retains all other copyrights.
Mostaghimi, Ali Reza, "Restriction Analysis of Bacteriophage" (1986). Loma Linda University Electronic Theses, Dissertations & Projects. 1440.
Loma Linda University Electronic Theses and Dissertations
Loma Linda University. Del E. Webb Memorial Library. University Archives