tag:blogger.com,1999:blog-32079676.post1312560342088226023..comments2024-02-13T21:22:02.522-08:00Comments on RRResearch: Reclaiming my explainer energy (from MOOCing back to blogging)Rosie Redfieldhttp://www.blogger.com/profile/06807912674127645263noreply@blogger.comBlogger2125tag:blogger.com,1999:blog-32079676.post-31773759408224896002013-07-19T18:04:22.524-07:002013-07-19T18:04:22.524-07:00We know from experiments I did 20 years ago that c...We know from experiments I did 20 years ago that crosslinking by mitomycin C doesn't induce competence. Now I think it's important that the fork stalling be due to nucleotide scarcity, so HU is just what we want.<br /><br />So I just did a Google Scholar search for 'hydroxyurea Escherichia coli' and found two nice papers from 1967 and 1972 that showed excellent inhibition, and the range of test conditions to use. Now I just need to check that these conditions aren't toxic to H. influenzae.Rosie Redfieldhttps://www.blogger.com/profile/06807912674127645263noreply@blogger.comtag:blogger.com,1999:blog-32079676.post-86935884620685241232013-07-18T01:40:09.977-07:002013-07-18T01:40:09.977-07:00You are right, hydroxyurea is the stuff to use to ...You are right, hydroxyurea is the stuff to use to induce replication fork stalling. Since you inhibit RNR, it is also reversible by washing your culture. I'm not really sure if bacterial RNR is affected by HU, though. I'm only aware of HU being used in eukaryotes.<br />Alternatively, you could try camptothecin that induces fork stalling by TOP1 inhibition. This is also reversible, but again I don't know about effectivity in bacteria.<br />An approach that will work in all organisms is cross-linking of DNA by Mitomycin C, but that is definitely irreversible.<br />In all of those cases you will also have to think about repair pathways, since fork stalling will induce homologous recombination.Alexanderhttp://scienceblogs.de/alles-was-lebt/noreply@blogger.com