Dave,

EcmA0-labile gfp gives signficantly better prestalk localization than
ecmA0 stable gfp.  The same applies for ecm0 constructs.  In both cases
the stable marker lights up many AL cells, while the labile marker is
confined to the canonical expression zone.

These constructs are now in the stock center, as user-friendly V18Tn5
vectors.  Transformant selection with these vectors is fast (5 days from
electroporation to clones with glowing slugs), simple (slugs form right on
the selection plates) and highly resistant to contamination (absolutely
no problem if the plates fall on the floor and open up).  You might thus
want to consider making the transformation part of the student lab
exercise.

Harry

-------------------------
On Mon, 6 Sep
2010, David Knecht charter wrote:

> We have found that the inexpensive LED illuminator system used in our teaching lab microscopes works fine for imaging cells in aggregates and slugs.  I would like to set up a new experiment for my course in which we image aggregation and later development using cell-type specific fluorescent protein probes to show the patterning of the slugs and fruiting bodies.  I intend to see if we can then do some slug surgery/mixing experiments to show sorting (mix the slug with a needle to start).  We can do this by mixing non-cell type specific labeled cells, but I would also like to use cell-type specific labels.   In Dictybase, I have found ecmA-GFP and cotB-GFP, but I haven't found any cell type specific red probes so that we can image two different cell type localizations in the same aggs/slugs.  Does anyone have a mRFP version of a cell type specific marker?
> Any suggestions for interesting variations on this experimental theme that can be done in a teaching lab are also welcome!
> THanks- Dave
>
>
> Dr. David Knecht
> Department of Molecular and Cell Biology
> Co-head Flow Cytometry and Confocal Microscopy Facility
> U-3125
> 91 N. Eagleville Rd.
> University of Connecticut
> Storrs, CT 06269
> 860-486-2200
> 860-486-4331 (fax)
>

----------------------------

Prof. Harry K. MacWilliams
Biologiezentrum der Ludwig-Maximilians-Universität
Grosshadenerstrasse 2
82152 Planegg-Martinsried
GERMANY

tel (office) +89 2180 74 288
tel (lab)    +89 2180 74 289
fax.         +89 2180 74 219

callers from North America please dial:
tel (office) 0114989 2180 74 288
tel (lab)    0114989 2180 74 289
fax          0114989 2180 74 219