Cite as: Cold Spring Harb. Protoc.; 2008; doi:10.1101/pdb.prot5102

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gaudet, P.
Right arrow Articles by Chisholm, R.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Gaudet, P.
Right arrow Articles by Chisholm, R.
Related Collections
Right arrow Cell Biology, general
Right arrow Laboratory Organisms, general
Right arrow Molecular Biology, general
Right arrow Developmental Biology
Right arrow Emerging Model Organisms
Right arrow DNA Delivery/Gene Transfer
Right arrow DNA Delivery/Gene Transfer, general
Right arrow Non-Viral Methods
Right arrowRelated Article
Right arrowRelated Protocols
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?
Legend icon

protocolProtocol

Transformation of Dictyostelium with Plasmid DNA by Calcium Phosphate Precipitation

Pascale Gaudet1, Petra Fey, and Rex Chisholm

dictyBase, Center for Genetic Medicine, Northwestern University, Chicago, IL 60611, USA

1Corresponding author (pgaudet{at}northwestern.edu)


INTRODUCTION

Dictyostelium discoideum is a unicellular eukaryote often referred to as a social ameba because it can form a multicellular structure when nutrient conditions are limiting. General principles for cell-to-cell communication, intracellular signaling, and cytoskeletal organization during cell motility have been derived from cellular and molecular studies of Dictyostelium and have been found to be conserved across all eukaryotes. The availability of a complete genome database and stocks of wild-type and mutant strains make D. discoideum an accessible and powerful model organism. Dictyostelium is amenable to genetic manipulations that require the introduction of DNA into cells, such as gene knockout, overexpression, antisense RNA expression, RNA interference (RNAi)-mediated gene knockdown, and restriction-enzyme-mediated mutagenesis. Calcium phosphate precipitation is a commonly used method for DNA-mediated transformation in Dictyostelium. Calcium phosphate precipitation produces high-copy-number transformants and is often used for overexpression experiments in conjunction with the G418 resistance gene, which needs to be present at high levels to produce efficient selection.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?

Related Article

Dictyostelium discoideum: The Social Ameba
Pascale Gaudet, Petra Fey, and Rex Chisholm
Cold Spring Harb Protoc 2008: 109. [Abstract] [Full Text]

Related Protocols

Growth and Maintenance of Dictyostelium Cells
Pascale Gaudet, Petra Fey, and Rex Chisholm
Cold Spring Harb Protoc 2008: 5099. [Abstract] [Full Text]

Making Permanent Stocks of Dictyostelium
Pascale Gaudet, Petra Fey, and Rex Chisholm
Cold Spring Harb Protoc 2008: 5101. [Abstract] [Full Text]

Transformation of Dictyostelium with Plasmid DNA by Electroporation
Pascale Gaudet, Petra Fey, and Rex Chisholm
Cold Spring Harb Protoc 2008: 5103. [Abstract] [Full Text]

Selection of Dictyostelium Transformants
Pascale Gaudet, Petra Fey, and Rex Chisholm
Cold Spring Harb Protoc 2008: 5104. [Abstract] [Full Text]



This article has been cited by other articles:


Home page
 Cold Spring Harb ProtocHome page
P. Gaudet, P. Fey, and R. Chisholm
Dictyostelium discoideum: The Social Ameba
Cold Spring Harb Protoc, December 1, 2008; 2008(13): pdb.emo109 - pdb.emo109.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
P. Gaudet, P. Fey, and R. Chisholm
Transformation of Dictyostelium with Plasmid DNA by Electroporation
Cold Spring Harb Protoc, December 1, 2008; 2008(13): pdb.prot5103 - pdb.prot5103.
[Abstract] [Full Text]

Home page
 Cold Spring Harb ProtocHome page
P. Gaudet, P. Fey, and R. Chisholm
Selection of Dictyostelium Transformants
Cold Spring Harb Protoc, December 1, 2008; 2008(13): pdb.prot5104 - pdb.prot5104.
[Abstract] [Full Text]