Skip to content Skip to navigation
University of Warwick
  • Study
  • |
  • Research
  • |
  • Business
  • |
  • Alumni
  • |
  • News
  • |
  • About

University of Warwick
Publications service & WRAP

Highlight your research

  • WRAP
    • Home
    • Search WRAP
    • Browse by Warwick Author
    • Browse WRAP by Year
    • Browse WRAP by Subject
    • Browse WRAP by Department
    • Browse WRAP by Funder
    • Browse Theses by Department
  • Publications Service
    • Home
    • Search Publications Service
    • Browse by Warwick Author
    • Browse Publications service by Year
    • Browse Publications service by Subject
    • Browse Publications service by Department
    • Browse Publications service by Funder
  • Help & Advice
University of Warwick

The Library

  • Login
  • Admin

Polynucleotide phosphorylases from thermophiles

Tools
- Tools
+ Tools

Wood, John N. C. (1976) Polynucleotide phosphorylases from thermophiles. PhD thesis, University of Warwick.

[img]
Preview
PDF
WRAP_Theses_Wood_1976.pdf - Unspecified Version - Requires a PDF viewer.

Download (5Mb) | Preview
Official URL: http://webcat.warwick.ac.uk/record=b1748203~S15

Request Changes to record.

Abstract

A purification procedure was developed for Eschericia coli polynucleotide phosphorylase, and subsequently applied to polynucleotide phosphorylases from Thermus aquaticus and Bacillus stearothermophilus. Preliminary investigations of the catalytic properties of the thermostable polynucleotide phosphorylases were carried out in the hope of effecting the facile polymerisation of modified nucleotide diphosphates which have a predominantly syn conformation. However, even at elevated temperatures, where the relative proportion of substrate molecules in the anti-conformation may be increased, the specificity of the thermostable enzymes was no broader than that reported for mesophylic enzymes. Other catalytic properties investigated were also similar to those observed using polynucleotide phosphorylases from other sources.

Structural studies of the enzyme from B. stearothermophilus revealed a similar gross amino acid composition and molecular weight to the E. coli enzyme. The quaternary structure differs from other polynucleotide phosphorylases in that four apparently identical subunits were identified on polyacrylamide gel electrophoresis under denaturing conditions. The subunits have a molecular weight of 51,000 daltons. Suberimidate cross-linking experiments confirmed a tetrameric structure for the native enzyme. Partially purified polynucleotide phosphorylase from T. aquaticus had a molecular weight of more than 400,000 daltons as judged by gel filtration.

Using a 3' exonuclease from Krebs ascites cells to degrade the rapidly labelled giant nuclear RNA from SV 40 transformed mouse cells, the location of virus specific sequences was investigated by hybridisation to purified SV 40 DNA. An apparent enrichment of virus sequences with increasing degradation of the RNA molecules suggests that virus sequences are absent at the 3’ end of giant nuclear RNA.

Item Type: Thesis or Dissertation (PhD)
Subjects: Q Science > QP Physiology
Q Science > QR Microbiology
Library of Congress Subject Headings (LCSH): Nucleic acids, Thermophilic microorganisms, Enzymes -- Analysis, Escherichia coli
Official Date: May 1976
Dates:
DateEvent
May 1976UNSPECIFIED
Institution: University of Warwick
Theses Department: Department of Chemistry
Thesis Type: PhD
Publication Status: Unpublished
Extent: [13], 202, [8] leaves : illustrations, charts, samples
Language: eng

Request changes or add full text files to a record

Repository staff actions (login required)

View Item View Item

Downloads

Downloads per month over past year

View more statistics

twitter

Email us: wrap@warwick.ac.uk
Contact Details
About Us