From owner-drosophila@net.bio.net Wed May 26 23:00:00 1993 Path: biosci!parc!decwrl!ames!elroy.jpl.nasa.gov!usc!howland.reston.ans.net!zaphod.mps.ohio-state.edu!math.ohio-state.edu!magnus.acs.ohio-state.edu!csn!ub!acsu.buffalo.edu!ubvmsb.cc.buffalo.edu!visahl From: visahl@ubvmsb.cc.buffalo.edu (A. Henry Latorella) Newsgroups: bionet.drosophila Subject: REQUEST: advice on embryo reporter genes Message-ID: Date: 27 May 93 20:51:00 GMT Sender: nntp@acsu.buffalo.edu Organization: University at Buffalo Lines: 86 News-Software: VAX/VMS VNEWS 1.41 Nntp-Posting-Host: ubvmsb.cc.buffalo.edu 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@uno.cc.geneseo.edu Cheers, * * * * * * * * * * * * * * * * * * * * * * * John Haynie * haynie@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 * .