Hello,
I have several questions which i would like to put to people in
various areas of research - please accept my apologies if you receive
this more than once.
Bacteriology...
We have transformed E. coli DH5a and HB101 using plasmid
pUV-BAD to express GFP and found that the cells are not uniform in length.
Can someone who has seen either or both of these strains - preferably after
they have beenstained in some way inform us as to whether they have these
morphologies if they have not been tinkered with!
When bacteria are DAPI stained some cells look orange under UV light - is
there a reason for this?
Protozoology...
When a protist ingests DTAF stained bacteria, what happens to the
stain? Is it and its' fluorescence degraded in the food vacuole or
does it end up in the medium when it is excreted?
GFP...
The transformed cells do not fluoresce uniformly - why is the protein
switched on at different rates amongst cells under the same
conditions? Is it because of new cells forming - therefore, do you have
to wait until all cells reach stationary phase before they will all
fluoresce at their maximum?
Will all the cells eventually fluoresce? Is it possible that some of the
cells will be non-fluorescing or is this impossible
since the plasmid gives ampilillin resistance and they have grown on
plates containing ampicillin.
All suggestions gratefully received...
Karen
