From owner-celegans@net.bio.net Sat Aug 31 23:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!news.sgi.com!cgl!jchou From: jchou@cgl.ucsf.edu (Joe Chou) Newsgroups: bionet.celegans Subject: Re: Identifying transgenic worms Date: 1 Sep 96 16:39:39 GMT Organization: UCSF Computer Graphics Lab Lines: 37 Message-ID: References: NNTP-Posting-Host: socrates.ucsf.edu bill.nuttley@utoronto.ca writes: >I am a new post-doc in Derek van der Kooy's lab in Toronto. One of my main >goals is to use cosmid rescue to find the genes that are mutated in the >associative learning mutants which have been isolated in the lab. My >problem is that the standard rol-6 marker for transformants will >interfere with the learning assays. >My first thought is to use GFP as a marker to ID the transgenic worms. >Has anybody been doing this? Does this allow rapid detection of >transformed animals? >Alternatively, I'm open to suggestions for other markers which do not >impair the worm's ability to move normally. Any and all suggestions would >be greatly appreciated. In the Bargmann Lab, we typically use a temperature-sensitive allele of lin-15 (n765ts) as a co-injection marker, screening for non-Muv F1's at 20 degrees. Although much more laborious, people in the lab have also used a GFP marker in cases where lin-15 has interacted with the gene of interest. The problem is that unless you have very high transmission frequencies, you will have to constantly confirm that the GFP is still present, and when you do assays (chemotaxis for us, learning for you), you will have to score only those animals that express GFP -- much more of a pain than just scoring non-Muv adults. Good luck, Joe Chou -- - Joe Chou (jchou@cgl.ucsf.edu) http://devbio-mac1.ucsf.edu/ PGP public key available by finger or at WWW page PGP FP [004C 5A68 CC2F DA20 3999 3355 0E8D 7B3F] .