Genetic Analysis of a Mammalian Chromosomal Origin of Replication

Abstract

The main goal of the research proposal was to develop an assay system for studying the specific genetic elements, if any, involved in the initiation of DNA replication in mammalian cells as outlined in Task 1 (development of assay system, see Appendix Statement of Work). The completion of this task is outlined in the annual summary from last year and led to the first peer-reviewed publication resulting from this research proposal (See Annual Summary, 2000, and attached reprint Altman and Fanning, 2001). Briefly, a competitive polymerase chain reaction (PCR)-based nascent strand abundance assay was used to demonstrate the ability of a small 5.8 kb fragment of DNA, containing the DHFR ori-beta initiation region (IR), to support efficient origin activity when integrated into random ectopic positions in the hamster chromosome (Altman and Fanning, 2001). In addition, the integrated ori-beta IR functioned with the same efficiency as the endogenous ori-beta in CHOK1 cells, suggesting that the 5.8 kb fragment was an acceptable candidate for mutational analysis of the ori-beta IR.

Open PDF

Document Details

Document Type
Technical Report
Publication Date
Aug 01, 2001
Accession Number
ADA400485

Entities

People

  • Amy L. Altman

Organizations

  • Vanderbilt University

Tags

Communities of Interest

  • Biomedical

DTIC Thesaurus Topics

  • Anti-Bacterial Agents
  • Biological Sciences
  • Biology
  • Breast Cancer
  • Cell Line
  • Cells
  • Chemical Reactions
  • Chromosomes
  • Eukaryotes
  • Fungi
  • Genetics
  • Materials
  • Molecular Biology
  • Polymerase Chain Reaction
  • Recognition
  • Tumor Cell Line
  • Two Dimensional

Fields of Study

  • Biology

Readers

  • Molecular Genetics
  • Technical Research and Report Writing.

Technology Areas

  • Biotechnology