Abstract
We have developed a transient expression system for the protist Physarum polycephalum based on firefly luciferase. We demonstrate the utility of this system for comparing the activities of different promoters in Physarum amoebae, and also for detecting genetic elements that affect the level of gene expression. This system is likely to facilitate improvements in the stable transformation of this organism.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alam J, Cook JL (1990) Reporter genes: application to the study of mammalian gene transcription. Anal Biochem 188:245–254
Blindt AB, Chainey AM, Dee J, Gull K (1986) Events in the amoebal-plasmodial transition of Physarum polycephalum studied by enrichment for committed cells. Protoplasma 132:149–159
Burland TG, Pallotta D, Tardif MC, Lemieux G, Dove WF (1991) Fission yeast promoter-probe vectors based on hygromycin resistance. Gene 100:241–245
Burland TG, Bailey J, Adam I, Mukhopadhyay MJ, Dove WF, Pallotta D (1992) Transient expression in Physarum of a chloroampenicol acetyltransferase gene under the control of actin gene promoters. Curr Genet 21:393–398
Burland TG, Bailey J, Pallotta D, Dove WF (1993) Stable selective, integrative DNA transformation in Physarum. Gene 132:207–212
Dee J, Foxon JI, Anderson RW (1989) Growth, development and genetic characteristics of Physarum polycephalum amoebae able to grow in liquid axenic medium. J Gen Microbiol 153:1567–1588
DiLella AG, Hope DA, Chen H, Trumbauer M, Schwartz RJ, Smith RG (1988) Utility of firefly luciferase as a reporter gene for promoter activity in transgenic mice. Nucleic Acids Res 16:4159
Hamelin M, Adam L, Lemieux G, Pallotta D (1988) Expression of the three unlinked isocoding actin genes of Physarum polycephalum. DNA 7:317–328
Howard PK, Ahern KG, Firtel RA (1988) Establishment of a transient expression system for Dictyostelium discoideum. Nucleic Acids Res 16:2613–2623
Jacobs WR, Barletta RG, Udani R, Chan J, Kalkut G, Sosne G, Kieser T, Sarkis GJ, Hatfull GF, Bloom BR (1993) Rapid assessment of drug susceptibilities of Mycobacterium tuberculosis by means of luciferase reporter phages. Science 260:819–822
Kondo T, Takahashi N, Muramatsu M (1992) The regulation of the murine Hox-2.5 gene expression during cell differentiation. Nucleic Acids Res 20:5729–5735
Millar AJ, Short SR, Chua N-H, Kay SA (1992) A novel circadian phenotype based on firefly luciferase expression in transgenic plants. Plant Cell 4:1075–1087
Newlon CS, Theis JF (1993) The structure and function of yeast ARS elements. Curr Opin Genes Dev 3:752–758
Roobol A, Pogson CI, Gull K (1980) In-vitro assembly of microtubule proteins from myxamoebae of Physarum polycephalum. Exp Cell Res 130:203–215
Roobol A, Wilcox M, Paul ECA, Gull K (1984) Identification of tubulin isoforms in the plasmodium of Physarum polycephalum by in-vitro microtubule assembly. Eur J Cell Biol 23:24–28
Showe MK, Williams DL, Showe IC (1992) Quantitation of transient gene expression after electroporation. Nucleic Acids Res 20:3153–3157
Wet JR de, Wood KV, Helinski DR, DeLuca M (1985) Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli. Proc Natl Acad Sci USA 82:7870–7873
Wet JR de, Wood KV, DeLuca M, Helinski DR, Subramani S (1987) Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol 7:725–737
Williams TM, Burlein TM, Ogden S, Kricka LJ, Kant JA (1989) Advantages of firefly luciferase as a reporter gene: application to the interleukin-2 gene promoter. Anal Biochem 176:28–32
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bailey, J., Benard, M. & Burland, T.G. A luciferase expression system for Physarum that facilitates analysis of regulatory elements. Curr Genet 26, 126–131 (1994). https://doi.org/10.1007/BF00313799
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00313799