REQUEST: advice on embryo reporter genes
A. Henry Latorella
visahl at ubvmsb.cc.buffalo.edu
Thu May 27 15:51:00 EST 1993
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
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
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
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
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>
* * * * * * * * * * * * * * * * * * * * * *
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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