Purification and characterization of a DNA polymerase gamma from human trophoblast tissue Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/cc08hj41c

Descriptions

Attribute NameValues
Creator
Abstract or Summary
  • DNA polymerase gamma has been purified over 67,500-fold from human trophoblast tissue. Purification was achieved through successive chromatographies on phosphocellulose, DEAE-cellulose, DNA-cellulose, and high pressure gel exclusion through Fractogel TSK HW55. This preparation yielded a specific activity of 81 units/mg of protein using the preferred template-primer, poly (A)-oligo (dT)₁₂₋₁₃. Analysis by SDS-polyacrylamide gel electrophoresis revealed a single polypeptide band with molecular weight 94,000 daltons, confirming the molecular weight estimate determined by gel exclusion. Sedimentation analysis in high ionic strength revealed an enzyme with molecular weight of 160-180,000 daltons, suggesting the enzyme exist as a dimer of the 94,000 dalton polypeptide. The enzyme exhibits optimal activity with poly (A)-oligo (dT)₁₂₋₁₈ in the presence of 10 mM KPO₄, 90 mM KC1 and 0.5 mM Mn²⁺. This demonstrates the first structural analysis of a DNA polymerase gamma from a human origin.
Resource Type
Date Available
Date Copyright
Date Issued
Degree Level
Degree Name
Degree Field
Degree Grantor
Commencement Year
Advisor
Academic Affiliation
Non-Academic Affiliation
Subject
Rights Statement
Peer Reviewed
Language
Digitization Specifications
  • File scanned at 300 ppi (Monochrome, 8-bit Grayscale) using ScandAll PRO 1.8.1 on a Fi-6670 in PDF format. CVista PdfCompressor 4.0 was used for pdf compression and textual OCR.
Replaces
Additional Information
  • description.provenance : Made available in DSpace on 2013-07-30T15:10:47Z (GMT). No. of bitstreams: 1 WongScottW1984.pdf: 812791 bytes, checksum: 1ccf40392a1eeb3c5371a6c9d1f10d48 (MD5) Previous issue date: 1983-08-18
  • description.provenance : Approved for entry into archive by Patricia Black(patricia.black@oregonstate.edu) on 2013-07-29T16:01:57Z (GMT) No. of bitstreams: 1 WongScottW1984.pdf: 812791 bytes, checksum: 1ccf40392a1eeb3c5371a6c9d1f10d48 (MD5)
  • description.provenance : Approved for entry into archive by Deborah Campbell(deborah.campbell@oregonstate.edu) on 2013-07-30T15:10:47Z (GMT) No. of bitstreams: 1 WongScottW1984.pdf: 812791 bytes, checksum: 1ccf40392a1eeb3c5371a6c9d1f10d48 (MD5)
  • description.provenance : Submitted by Katy Davis (kdscannerosu@gmail.com) on 2013-07-25T18:03:39Z No. of bitstreams: 1 WongScottW1984.pdf: 812791 bytes, checksum: 1ccf40392a1eeb3c5371a6c9d1f10d48 (MD5)

Relationships

Parents:

This work has no parents.

Last modified

Downloadable Content

Download PDF

Items