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

Some studies on the use of polynucleotide phosphorylase in the synthesis of polynucleotides

Tools
- Tools
+ Tools

Smith, John C. (1975) Some studies on the use of polynucleotide phosphorylase in the synthesis of polynucleotides. PhD thesis, University of Warwick.

[img]
Preview
PDF
WRAP_Theses_Smith_1975.pdf - Unspecified Version - Requires a PDF viewer.

Download (8Mb) | Preview
Official URL: http://webcat.warwick.ac.uk/record=b1747688~S15

Request Changes to record.

Abstract

The purification of polynucleotide phosphorylase(E.C.2. 7. 7. 8) by affinity chromatography was investigated. Several affinity ligands were prepared and one of them, oligo(dT) Sepharose, was found to be an efficient adsorbent for the enzyme.
Polynucleotide phosphorylase purified from Micrococcusluteus and Escherichia coli, was immobilized by covalent attachment to a number of insoluble supports. The properties of the immobilized enzyme were compared with those of the enzyme in free solution. It was found that while there was no significant change in the kinetic parameters of the enzyme, there was a significant increase in the stability of the immobilized derivatives. Studies were made of the use of the immobilized enzyme in the synthesis of polynucleotides.
Polynucleotide phosphorylase was isolated from Bacillus Stearothermophilus and was used to incorporate CDP residues into long oligo nucleotide primers such as oligoi(1))and t-RNA. Primers were isolated by chromatography on dihydroxyboryl Sepharoso which ensured that the primers possessed an intact 3'-hydroxyl group. Although incorporation of CDP into the primer was achieved, the number of residues incorporated was low. However, the method could be used to label the 3-terminus of a polynucleotide, in an Appendix, file purification of an endonuclease from pig liver nuclei is described. The enzyme was used in the preparation of oligonucleotide primers.

Item Type: Thesis or Dissertation (PhD)
Subjects: Q Science > QD Chemistry
Library of Congress Subject Headings (LCSH): Nucleosides -- Synthesis, Nucleic acids -- Synthesis, Nucleotides, RNA polymerases
Official Date: November 1975
Dates:
DateEvent
November 1975Submitted
Institution: University of Warwick
Theses Department: Department of Chemistry
Thesis Type: PhD
Publication Status: Unpublished
Supervisor(s)/Advisor: Hutchinson, D. W. (David Wesley)
Sponsors: Science Research Council (Great Britain) ; Searle Research Laboratories
Extent: 78 leaves : illustrations
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