Identifying transgenic worms

Joe Chou jchou at cgl.ucsf.edu
Sun Sep 1 11:39:39 EST 1996


bill.nuttley at 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

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- Joe Chou  (jchou at cgl.ucsf.edu)
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