Restriction Enzyme Analysis of Incomplete Parental Replicative Forms of øX174

<p>The initial step in the infectious cycle of the single-stranded DNA phage øX174 is the synthesis of a DNA strand complementary to the infecting viral DNA producing a duplex molecule, the parental replicative form (RF). Electron micrographic examination of parental RF produced from viruses exposed to ultra-violet (UV) light suggests that synthesis begins at only one site. However, <em>in vitro</em> results and some recent reports of <em>in vivo</em> experiements contradict that interpretation.</p> <p>Using restriction enzymes we have re-examined the incomplete RF made from UV-damaged phage. Parental RF made <em>in vivo</em> from viruses exposed to various UV doses were purified by velocity sedimentation and benzoylated naphthoylated diethylaminoethylcellulose column chromatography. The incomplete parental RF were digested with a restriction enzyme and the fragments separated by by gel electrophoresis. The genetic location of the new DNA was determined from the relative amount of label appearing at each fragment position.</p> <p>Readioactive label was not uniformly distributed among the DNA fragments, but neither was there a simple sequence of increasing radioactivity. We conclude that initiation was not random but occurred at several sites. The data do not exclude the possibility that a single initiation site was employed on each template but that the site varied from template to template.</p>

Name thesis:degree_name

Master of Arts (MA)

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Biology

Year

1979

Author dc:creator

• Bogardus, Alicia Morales

Contributors dc:contributor

• Anthony J. Zuccarelli
• Leonard R. Brand
• Robert L. Nutter

• Biology
• Bacteriophages -- analysis; DNA Restriction Enzymes; DNA Single-Stranded

Statement dc:rights

• 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.

Language dc:language

English

Repository record dc:identifier

https://scholarsrepository.llu.edu/etd/1396

OAI identifier oai:identifier

oai:scholarsrepository.llu.edu:etd-2164