Transformation of Dictyostelium discoideum with plasmid DNA.

Transformation of Dictyostelium discoideum with plasmid DNA. Research Abstract Details 

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Transformation of Dictyostelium discoideum with plasmid DNA. Abstract Text:

Pascale Gaudet,Karen E Pilcher,Petra Fey,Rex L Chisholm,Pascale Gaudet,Karen E Pilcher,Petra Fey,Rex L Chisholm,

DNA-mediated transformation is one of the most widely used techniques to study gene function. The eukaryote Dictyostelium discoideum is amenable to numerous genetic manipulations that require insertion of foreign DNA into cells. Here we describe two commonly used methods to transform Dictyostelium cells: calcium phosphate precipitation, resulting in high copy number transformants; and electroporation, an effective technique for producing single integration events into genomic DNA. Single integrations are required for gene disruption by homologous recombination. We also discuss how different selection markers affect vector copy number in transformants and explain why blasticidin has become the preferred selectable marker for making gene knockouts. Both procedures can be accomplished in less than 2 h of hands-on time; however, the calcium phosphate precipitation method contains several incubations, including one of at least 4 h, so the total time required for the transformation is approximately 8 h.

Transformation of Dictyostelium discoideum with plasmid DNA. Publishing Authors By Initials

P Gaudet,KE Pilcher,P Fey,RL Chisholm,P Gaudet,KE Pilcher,P Fey,RL Chisholm,

For similar genetic processes: recombination, genetic: transformation, genetic research abstracts see: genetic processes: recombination, genetic: transformation, genetic research

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Transformation of Dictyostelium discoideum with plasmid DNA. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Nature protocols

VOLUME: 2

Page Numbers: 1317-24

Journal Abbreviation:

ISSN: 1750-2799

DAY: 3

MONTH: 12

YEAR: 2007

Transformation of Dictyostelium discoideum with plasmid DNA. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 101284307

Transformation of Dictyostelium discoideum with plasmid DNA. Keywords Mesh Terms:

KEYWORDS: Transformation, Genetic

MESH TERMS: genetics

Chemical & Substance for Abstract: Transformation of Dictyostelium discoideum with plasmid DNA. Information

Substance Name: DNA

Registry Number: 9007-49-2

Grant and Affiliation Information for Transformation of Dictyostelium discoideum with plasmid DNA.

AFFILIATION: dictyBase, Center for Genetic Medicine, Northwestern University, 676 North Saint Clair Street Suite 1260, Chicago, Illinois 60611, USA.

Country: England

AGENCY: United States NHGRI

GRANT: HG00022

ACRONYM: HG

MEDLINETA: Nat Protoc

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