Genetic interaction studies in bacteria are quite challenging. Before this study, we had data for hundreds of interactions in E. coli in comparison with thousands known for yeast. In this paper, the authors introduce a method termed GIANT which relies on massive Hfr conversion and double mutant generation in E. coli. The method is presented below as a figure from the original paper:
Here, we rely on conjugation to select for double markers (double mutants) and assay they growth on 384 or 1536 colony arrays. The authors make the case for their method through several validation steps. In the end, this method has the ability to generalize to other organisms for which deletion collections are available.
2 comments:
this is good way for studying intraction between some genes in E.Coli, but why is it difficult to study intactions in bacteria in camparision with yeast?
The most important reason used be the simplicity of generating deletion libraries in yeast compared to bacteria. However, the method of Datsenko and Wanner to a large extent revolutionized DNA manipulation in E. coli.
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