REQUEST: advice on embryo reporter genes

A. Henry Latorella visahl at ubvmsb.cc.buffalo.edu
Thu May 27 15:51:00 EST 1993


INTRO:
Next year I plan to introduce the ideas of reporter genes 
and enhancer traps into my undergraduate (junior/senior) 
developmental biology teaching laboratory.  I'm seeking the 
advice of this group in order to get started on the right 
foot.  

BACKGROUND:
Students in past years have succeeded in looking at live and 
fixed/Feulgen-stained embryos; I have no experience with 
beta-gal staining of Drosophila embryos but from the 
Ashburner book, I'm guessing that it won't add that much 
difficulty to the protocol.  I don't believe that in situ 
hybridization is feasible under the circumstances (number 
and expertise of students) but let someone out there 
convince me that I am wrong.

MY NAIVE IDEA:

This is what I'd like to do in general:
                           ^^^^^^^^^^^
1) Obtain one or two strains (made by jumping or by 
construct/transformation) which  express beta-gal in an 
interesting temporal/spatial pattern (perhaps a gap gene and 
a pair-rule gene).  Students can make timed collections and 
stain with X-gal to see this pattern.

2) (optional) Cross these reporters into other backgrounds 
to demonstrate modifications of their pattern of expression.  
This could include maternal effect as well as other zygotic 
segmentation loci.

Specifically:
^^^^^^^^^^^^^
I know that there are ftz/lac-Z constructs available from 
IUBIO stock center (although I haven't figured out which is 
which).  I'm assuming that the "reporter" gene is not 
segregating in these stocks.  These will presumably give the 
archetypal 7 stripes.  Just about any issue of CELL has 
references to other strains which I'm assuming I could 
obtain from their creators for my stated purpose.  (Which of 
these will give the best results considering that this is 
mostly for demonstration purposes?)

Students can measure the %EggLength of each of the "ftz" 
stripes and do the same in embryos which have been laid by 
mothers with varying doses of bcd+.  (There must be a large 
number of "interesting" combinations of defective 
genotype/reporter gene to illustrate the phenomenon of 
interaction.)

MY REQUEST:

I'd like to have feedback from anyone with practical 
experience in this kind of protocol so that I can identify 
and obtain strains with the best practical characteristics 
(strong expression, stable stocks, etc.).  

Any suggestions about good combinations of 
regulators/reportors will also be much appreciated.

Finally, if you can point me to a source of this 
information, I will thank you very much!

WANT A SUMMARY?

If there is clear interest that I do so, I can post a 
summary.  Otherwise, I can e-mail what I collect to 
interested individuals.

Please reply directly at the address give below.  I am 
reading the news as a guest on another machine.

haynie at uno.cc.geneseo.edu		<John Haynie>

Cheers,


*  *  *  *  *  *  *  *  *  *  *  *  *  *  *  *  *  *  *  *  *  *
				*
John Haynie			*	haynie at uno.cc.geneseo.edu
Biology Department		*	tel: 716) 245-5306
State University of New York	*	fax: 716) 245-5007
College of Arts and Sciences	*
Geneseo, NY 14454		*



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