Literature DB >> 9007012

Wild-type strains of Dictyostelium discoideum can be transformed using a novel selection cassette driven by the promoter of the ribosomal V18 gene.

B Wetterauer1, P Morandini, I Hribar, I Murgia-Morandini, U Hamker, C Singleton, H K Macwilliams.   

Abstract

Almost all methods for transformation of the social ameba Dictyostelium discoideum rely on axenic growth, that is, growth in a synthetic medium, for at least part of the procedure. Axenic growth requires several mutations. Here we describe a procedure that can be used to transform wild-type strains which are able to grow only on the natural food source, bacteria. The method relies on a new selection cassette driven by the V18 promoter, a promoter that we show is substantially more active during growth on bacteria than the actin-6 promoter, which is widely used for axenic transformation. The procedure gives transformation frequencies of about 10(-5) with both strains Ax2 (capable of axenic growth) and NC4 (capable of growth only on bacteria). Using this vector, we have obtained NC4 strains carrying several beta-galactosidase reporter cassettes. Our vector can also be used in axenic transformations.

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Year:  1996        PMID: 9007012     DOI: 10.1006/plas.1996.0044

Source DB:  PubMed          Journal:  Plasmid        ISSN: 0147-619X            Impact factor:   3.466


  7 in total

1.  Multiple signalling pathways connect chemoattractant receptors and calcium channels in Dictyostelium.

Authors:  Thomas Nebl; Martha Kotsifas; Pauline Schaap; Paul R Fisher
Journal:  J Muscle Res Cell Motil       Date:  2002       Impact factor: 2.698

2.  Involvement of a novel gene, zyg1, in zygote formation of Dictyostelium mucoroides.

Authors:  Aiko Amagai
Journal:  J Muscle Res Cell Motil       Date:  2002       Impact factor: 2.698

3.  Inducible expression of exogenous genes in Dictyostelium discoideum using the ribonucleotide reductase promoter.

Authors:  P Gaudet; H MacWilliams; A Tsang
Journal:  Nucleic Acids Res       Date:  2001-01-15       Impact factor: 16.971

4.  A cysteine-rich extracellular protein containing a PA14 domain mediates quorum sensing in Dictyostelium discoideum.

Authors:  Alexandra Kolbinger; Tong Gao; Debbie Brock; Robin Ammann; Axel Kisters; Joseph Kellermann; Diane Hatton; Richard H Gomer; Birgit Wetterauer
Journal:  Eukaryot Cell       Date:  2005-06

5.  Rapid and efficient genetic engineering of both wild type and axenic strains of Dictyostelium discoideum.

Authors:  Peggy Paschke; David A Knecht; Augustinas Silale; David Traynor; Thomas D Williams; Peter A Thomason; Robert H Insall; Jonathan R Chubb; Robert R Kay; Douwe M Veltman
Journal:  PLoS One       Date:  2018-05-30       Impact factor: 3.240

6.  Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria.

Authors:  Peggy Paschke; David A Knecht; Thomas D Williams; Peter A Thomason; Robert H Insall; Jonathan R Chubb; Robert R Kay; Douwe M Veltman
Journal:  J Vis Exp       Date:  2019-01-25       Impact factor: 1.355

7.  Replacement of the essential Dictyostelium Arp2 gene by its Entamoeba homologue using parasexual genetics.

Authors:  Mehreen Zaki; Jason King; Klaus Fütterer; Robert H Insall
Journal:  BMC Genet       Date:  2007-06-06       Impact factor: 2.797

  7 in total

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