skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Homologous gene replacement in Physarum

Abstract

The protist Physarum polycephalum is useful for analysis of several aspects of cellular and developmental biology. To expand the opportunities for experimental analysis of this organism, we have developed a method for gene replacement. We transformed Physarum amoebae with plasmid DNA carrying a mutant allele, ardD{Delta}1, of the ardD actin gene; ardD{Delta}1 mutates the critical carboxy-terminal region of the gene product. Because ardD is not expressed in the amoeba, replacement of ardD{sup +} with ardD{Delta}1 should not be lethal for this cell type. Transformants were obtained only when linear plasmid DNA was used. Most transformants carried one copy of ardD{Delta}1 in addition to ardD{sup +}, but in two (5%), ardD{sup +} was replaced by a single copy of ardD{Delta}1. This is the first example of homologous gene replacement in Physarum. ardD{Delta}1 was stably maintained in the genome through growth, development and meiosis. We found no effect of ardD{Delta}l on viability, growth, or development of any of the various cell types of Physarum. Thus, the carboxy-terminal region of the ardD product appears not to perform a unique essential role in growth or development. Nevertheless, this method for homologous gene replacement can be applied to analyze the function of any cloned gene.more » 38 refs., 6 figs., 1 tab.« less

Authors:
 [1];  [2]
  1. Univ. of Wisconsin, Madison, WI (United States)
  2. Laval Univ., Quebec (Canada)
Publication Date:
OSTI Identifier:
255334
Resource Type:
Journal Article
Resource Relation:
Journal Name: Genetics; Journal Volume: 139; Journal Issue: 1; Other Information: PBD: Jan 1995
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; GENES; GENE MUTATIONS; STRUCTURE-ACTIVITY RELATIONSHIPS; THERAPY; GENETIC MAPPING; TRANSCRIPTION; PHYSARUM; GROWTH; DNA-CLONING; POLYMERASE CHAIN REACTION; DNA SEQUENCING

Citation Formats

Burland, T.G., and Pallotta, D.. Homologous gene replacement in Physarum. United States: N. p., 1995. Web.
Burland, T.G., & Pallotta, D.. Homologous gene replacement in Physarum. United States.
Burland, T.G., and Pallotta, D.. Sun . "Homologous gene replacement in Physarum". United States.
@article{osti_255334,
title = {Homologous gene replacement in Physarum},
author = {Burland, T.G. and Pallotta, D.},
abstractNote = {The protist Physarum polycephalum is useful for analysis of several aspects of cellular and developmental biology. To expand the opportunities for experimental analysis of this organism, we have developed a method for gene replacement. We transformed Physarum amoebae with plasmid DNA carrying a mutant allele, ardD{Delta}1, of the ardD actin gene; ardD{Delta}1 mutates the critical carboxy-terminal region of the gene product. Because ardD is not expressed in the amoeba, replacement of ardD{sup +} with ardD{Delta}1 should not be lethal for this cell type. Transformants were obtained only when linear plasmid DNA was used. Most transformants carried one copy of ardD{Delta}1 in addition to ardD{sup +}, but in two (5%), ardD{sup +} was replaced by a single copy of ardD{Delta}1. This is the first example of homologous gene replacement in Physarum. ardD{Delta}1 was stably maintained in the genome through growth, development and meiosis. We found no effect of ardD{Delta}l on viability, growth, or development of any of the various cell types of Physarum. Thus, the carboxy-terminal region of the ardD product appears not to perform a unique essential role in growth or development. Nevertheless, this method for homologous gene replacement can be applied to analyze the function of any cloned gene. 38 refs., 6 figs., 1 tab.},
doi = {},
journal = {Genetics},
number = 1,
volume = 139,
place = {United States},
year = {Sun Jan 01 00:00:00 EST 1995},
month = {Sun Jan 01 00:00:00 EST 1995}
}