Hi,
Does any one have a strain with gfp driven by an osm-6 promoter that has been integrated and outcrossed? Or is it available on CGC?
Please help me out!
Regards
Sudhanva
Hi,
Does any one have a strain with gfp driven by an osm-6 promoter that has been integrated and outcrossed? Or is it available on CGC?
Please help me out!
Regards
Sudhanva
Hi,
have you tried looking at the entry for osm-6 in wormbase, specifically the experimental evidence for expression using the osm-6::gfp construct?
or here;
http://www.shigen.nig.ac.jp/c.elegans/promoter/DetailSearch?no=90&lang=english
or perhaps contact this group who also used this construct;
http://www.nature.com/ncb/journal/v14/n9/full/ncb2560.html
Regards
Steve
Hi Steve,
Thanks for that! I did read that paper. I think they have used an OSM-6-GFP fusion. Where as I need gfp expressed under an osm-6 promoter as I want to look at degeneration in sensory neurons. I’d prefer wild-type expression of osm-6 in my worms.
Regards
Sudhanva
Hi Sudhanva,
in the absence of any further info regarding an integrated osm-6promoter::gfp strain, I guess you are in the same position as me and will have to do the transgenics yourself. I’m presently setting this up in my lab.
Regards
Steve
Sudhanva,
Did you try following Steve’s earlier advice, and looking at the WormBase page for osm-6? Specifically, in the Reagents box?
The table isn’t displaying allele names correctly for some reason, but if you click on the transgenes you’ll find at least five that claim to be fairly straightforward integrated osm-6::gfp transgenes (albeit a couple of them with a Rol phenotype): cgc7279Is1, lqIs2, lqIs3, minIs12, and oyIs59. The first has an odd name and no further information, but it should be straightforward to seek out more information on the others, for example by contacting the originating labs. It is of course possible that some or all of these express a GFP fusion to OSM-6 (“osm-6::gfp” doesn’t really distinguish transcriptional from translational fusions), but it’s likely some do not, and simply express GFP under the osm-6 promoter, which I take to be what you’re looking for.
Thanks HillelSchwartz! I didn’t expect them to be in the Reagents box!
Steve, looks like Eric Lundkvist’s lab at the University of Kansas, USA has the strain we’re looking for! Considering they have made, integrated and mapped the integration on to the 4th chromosome, I am pretty sure they must have outcrossed it as well. I am sending them an email asking for a plate! (I am not too sure if you meant Posm-6::gfp when you said you’re setting ‘this’ up in your lab)
Regards
Sudhanva
Hi,
yes, that was a good tip from Hillel.
No I’m not using Posm-6 for my transgenics, I’m designing some transgenes with different promoter-receptor-gfp fusions…unfortunately (or perhaps not) there are no existing strains so I have to make my own…the winters are sometimes long here so there’s plenty of time.
Anyway, glad you have found a possible source for the transgene…is it only held by Eric and not by the CGC?
Regards
Steve