vital GUS staining - summary

Ed Rybicki ed at micro.uct.ac.za
Tue May 24 10:43:26 EST 1994


Summary of responses to the request for methods for staining 
tissue for GUS without killing the tissue: many thanks to all those who 
unselfishly shared all their tips and/or references; you were a great help.

Three Plant Molecular Biology Reporter articles give methods for 
non destructive GUS assays:

J.H. Gould and R.H. Smith (1989).  A non-destructive assay for 
GUS in the media of plant tissue cultures.  Plant Molecular 
Biology Reporter 7(3): 209-216.

Martin et al. Non-destructive assay system for detection of GUS 
activity in higher plants.  Plant Molecular Biology Reporter 
10(1): 37-46.

G. Kirchner, C.J. Kinslow, G.C. Bloom and D.W. Taylor. (1993).  
Nonlethal assay system of B-Glucuronidase activity in transgenic 
tobacco roots.  Plant Molecular Biology Reporter 11(4): 320-325.

In addition, Richard Jefferson comments in his Plant Molecular 
Biology Reporter article (1987; vol5 no 4, p399) that 
"reasonable staining can be achieved at much lower substrate 
concentrations (50ug/ml) with enhanced recovery (of stained 
cells)."  

Also, in a recent article, Swoboda et al. (1994) EMBO J. 13: 
484-489 have an in vivo staining procedure they use on 
Arabidopsis rosettes:
Vacuum infiltrate for 5 minutes with sterile staining buffer 
containing 200 mg X-Gluc in 300 ml 50 mM phosphate buffer (pH 
7.0) 0.5 x MS medium.  Incubate 4-8 hours at 37 degrees.

Martin et al. in Sean Gallager's book "Gus Protocols" - Academic 
Press 1992 has a method for using MUG in intact tissues.  We we 
have used this technique to confirm that seedlings were 
transgenic and it seems to work okay.  We did not try to 
continue growing the plants, but the authors suggest it is 
possible (Prof. David Oliver , University of Idaho)


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