Abstract

Over 90% recovery of phage P4/2 was realized with differential centrifugation concentration and purification, whereas only about 16% was recovered from the aqueous two-phase polymer - Genetron - ECTEOLA-cellulose column system. Purity of preparations was greatest with the density gradient centrifugation (A260 = 2.66 and A280 = 1.80 per 1012 plaque-forming units). With the chemical system of concentration and purification followed by an ECTEOLA anion exchange run, the A260 was 6.6 and the A280 was 4.3.

Three tests were employed to determine the configuration of P4/2 DNA: (1) a formalin denaturation test, (2) a thermal gradient analysis, and (3) a heat denaturation test. All indicated a double-stranded nature.

Eight determinations each for the amounts of DNA, phosphorus and protein present in P4/2 showed 1.26 x 10-16 gm, 1.98 x 10-17 gm, and 1.63 x 10-16 gm per pfu, respectively. Since the sample under analysis had been prepared by the chemical method, which involved a Waring blender treatment, a factor was used to correct for inactivated phages present. The factor was based on the calculated weight of the phage from the density of the particles in equilibrium sedimentation multiplied by the volume of the particles estimated from electron micrographs. This was expressed as a ratio over the chemically determined weight of the phage. The revised values were: DNA 0.80 x 10-16 gm, phosphorus 1.25 x 10-17 gm, and protein 1.03 x 10-16 gm per particle, giving phage P4/2 chemical values similar to those of phage T7 with which it is quite similar morphologically.

Five descending chromatography determinations were performed on hydrolyzed P4/2 DNA yielding the following base ratio averages: guanine 23.1%, adenine 27.4%, cytosine 23.4%, and thymine 26.3%. These values compare with the base ratios of the host bacterial DNA within probable experimental error.

LLU Discipline

Biology

Department

Biology

School

Graduate School

First Advisor

Robert L. Nutter

Second Advisor

Leonard R. Bullas

Third Advisor

Norman M. Case

Fourth Advisor

Earl W. Lathrop

Fifth Advisor

Ariel A. Roth

Degree Name

Master of Arts (MA)

Degree Level

M.A.

Year Degree Awarded

1966

Date (Title Page)

9-1966

Language

English

Library of Congress/MESH Subject Headings

Salmonella

Type

Thesis

Page Count

viii; 49

Digital Format

PDF

Digital Publisher

Loma Linda University Libraries

Usage 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.

Collection

Loma Linda University Electronic Theses and Dissertations

Collection Website

http://scholarsrepository.llu.edu/etd/

Repository

Loma Linda University. Del E. Webb Memorial Library. University Archives

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