hfq results

Yes indeed, hfq is needed for full competence development.  The mutant grows as well as its wild-type parent (top graph) but develops about ten-fold lower competence (lower graph).

 

Next step:  Make DNA from the mutant and use it to transform the hfq knockout into the hypercompetent mutants.  Then I'll test the effect of the hfq knockout on their competence.  If Hfq avcts directly on the sxy mRNA stem that regulates translation, I expect the mutants to be unaffected by loss of Hfq because their translation is not limited by the stem.  

As a check that Hfq's effects aren't due to indirect effects on cAMP levels or CRP activity, I'll also test the effect of the knockout in wildtype and hypercompetent cells with added cAMP.

What about effects on the murE hypercompetence mutants?  I'll test that, but I'm not sure how to interpret different results...