gene replacements can be performed in species, like yeast and bacteria, that exh
ID: 62797 • Letter: G
Question
gene replacements can be performed in species, like yeast and bacteria, that exhibit high rates of homologous recombination (knockouts are just one example of this). Suppose that someone hands you a 3 kb BamHI fragment of DNA that they say encodes a Pseudomonas sigma factor. Describe an experiment that you could perform to test whether that gene could replace the sigma 70 gene in E. coli, i.e. it will work like a normal sigma factor in E. coli. As part of your answer, describe the detailed features of any DNA construct that you will need to make, as well as the general strategy of your experiment.
Explanation / Answer
Take a sigma 70 knockout E.coli, and transform the 3 kb BamH1 fragment into the E.coli. If the Pseudomonas sigma factor could replace the sigma 70 gene, then the E.coli survives and shows growth on the medium, otherwise the bacteria would die.
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.