Optimization of a large-scale gene disruption protocol in Dictyostelium and analysis of conserved genes of unknown function

BACKGROUND: Development of the post-genomic age in Dictyostelium will require the existence of rapid and reliable methods to disrupt genes that would allow the analysis of entire gene families and perhaps the possibility to undertake the complete knock-out analysis of all the protein-coding genes present in Dictyostelium genome. RESULTS: Here we present an optimized protocol based on the previously described construction of gene disruption vectors by in vitro transposition. Our method allows a rapid selection of the construct by a simple PCR approach and subsequent sequencing. Disruption constructs were amplified by PCR and the products were directly transformed in Dictyostelium cells. The selection of homologous recombination events was also performed by PCR. We have constructed 41 disruption vectors to target genes of unknown function, highly conserved between Dictyostelium and human, but absent from the genomes of S. cerevisiae and S. pombe. 28 genes were successfully disrupted. CONCLUSION: This is the first step towards the understanding of the function of these conserved genes and exemplifies the easiness to undertake large-scale disruption analysis in Dictyostelium

Mono, mixed or multiple strategy approach: a descriptive study of the latest published articles in the International Journal of Project Management

To develop a valid and credible research project, it is essential to define in detail the research design. It will have a meaningful strategy to answer the research question. While some researchers suggest that there might be some benefits in adopting multiple and mixed strategy approach for conducting the research, some tend to stick to mono-methods. This article tackles a descriptive study that identifies the research strategies followed by top ranked researches, by reviewing the last two years of publications of the International Journal of Project Management, the leading journal for the field of project management and organization studies. The results suggest that most researchers stick to mono-methods, predominantly with qualitative techniques. Also, there is a higher trend to use case study strategies in the field