tag:blogger.com,1999:blog-32079676.post1397933775351459917..comments2024-02-13T21:22:02.522-08:00Comments on RRResearch: Why is H. influenzae's CRP so feeble?Rosie Redfieldhttp://www.blogger.com/profile/06807912674127645263noreply@blogger.comBlogger4125tag:blogger.com,1999:blog-32079676.post-70651514772480903362007-02-15T20:41:00.000-08:002007-02-15T20:41:00.000-08:00Did you guys try an overexpression of yccR(sxy) an...Did you guys try an overexpression of yccR(sxy) and see how it would effect E. coli's comF(yhgH) mRNA levels? Because yrfD is induced and it got two CRP-S sites, why not comF? Also, how about indution of other genes in the yrfD operon suc as yrfC, yrfB, yrfA and hofQ? However, I expected to be induced as well but would it be at the same level as yrfD mRNA transcripts?Slavickhttps://www.blogger.com/profile/17282372934696400020noreply@blogger.comtag:blogger.com,1999:blog-32079676.post-18416081874467813402007-02-09T11:58:00.000-08:002007-02-09T11:58:00.000-08:00Heather, Andrew agrees with your idea.Keith, yes, ...Heather, Andrew agrees with your idea.<BR/><BR/>Keith, yes, the affinity measurements were done using band-shifts in vitro. We knwo that E. coli CRP works fine in H. influenzae, but we haven't yet looked at kinetics of induction of CRP-dependent genes. One simple experiment would be to find out how much cAMP is needed to induce the genes when the different CRPs are present.Rosie Redfieldhttps://www.blogger.com/profile/06807912674127645263noreply@blogger.comtag:blogger.com,1999:blog-32079676.post-37236190392047296762007-02-08T21:18:00.000-08:002007-02-08T21:18:00.000-08:00I'm assuming from the way you describe things that...I'm assuming from the way you describe things that these are in vitro affinity measurements -- is that correct?<BR/><BR/>What happens if you replace E.coli CRP with H.influenzae CRP? Does it change when catabolite repression turns on & off?Keith Robisonhttps://www.blogger.com/profile/04765318239070312590noreply@blogger.comtag:blogger.com,1999:blog-32079676.post-55139794609095004342007-02-08T18:41:00.000-08:002007-02-08T18:41:00.000-08:00I think it has to do with a discussion we had earl...I think it has to do with a discussion we had earlier this year, that H. influenzae is "on its way" to having a reduced genome. There probably isn't as much purifying selection acting on CRP in Hin, due to fewer genes in its regulon for example, and therefore deleterious mutations are not purged as readily as they are in E. coli.90% microbehttps://www.blogger.com/profile/10288595333674587852noreply@blogger.com