DICTYOSTELIUM DISCOIDEUM

Recommendations

Recent advances in microbial and molecular genetics in D. discoideum have permitted rapid construction of complex genotypes. Moreover, detailed maps of the 6 chromosomes (resolution 50 kb) and ordered sets of YACs representing the 34 Mb genome are available. The complete genome is being sequenced. An EST project in Japan has sequenced 10 000 cDNAs from mid-development stages and is in the process of sequencing another 10 000 cDNAs from growing cells.

Efficient homologous recombination permits rapid gene replacement. Since D. discoideum grows and develops equally well as a haploid or a diploid, it can be rapidly screened for various phenotypes, including defects in phagocytosis, cytokinesis, cell motility, chemotaxis, and other cell-cell signaling networks. Random insertional mutagenesis has allowed over 150 developmentally essential genes to be characterized. The frequency of mutagenesis is sufficiently high to permit saturation suppression screens. Because development is gratuitous mutants strains can be grown and stored in a reliable manner. High resolution computer assisted microscopy of pertinent mutant strains has led to sophisticated phenotypic analysis.

NEEDS OF THE COMMUNITY

  1. Completion of the genome sequence (finishing steps). Approximately 3 million dollars.
  2. Expansion of existing AceDB databases of genomic, mutational, and analytic information. Approximately 200 000 dollars/year.
  3. Organization of a centralized stock storage and distribution center for plasmids and mutant strains. Approximately 300 000 dollars/year.

RESOURCES FOR THE FUTURE

  1. Arrayed DNA on chips to permit expression studies of every ORF throughout the 24 hour developmental cycle.
  2. Proteome studies - patterns of protein accumulation in wild type and mutant strains throughout the 24 hour developmental cycle.
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