Fire Lab Vector Kit terminology

Model Organism: Worms Methods & Reagents
1

Does anyone know where I can find a guide to deciphering the various descriptions of the Fire Lab Vectors (e.g. dpy30Tsa/gfpN/IVSR/gfpS65C/IVSS/IVST/gfpC/let8583’/decoy/)? I’m particularly worried about the “decoy”, bit, which I just noticed.
I attempted to make a translational fusion using the ppD152.79 vector (L5494, http://www.addgene.org/1704/ ), and it looks in frame and correct by sequencing, but I haven’t been able to generate any transgenics by injection (no problem with the empty vector though). This has occurred for 2 different fusion proteins, so I don’t think it’s the actual genes themselves giving me the problem. If you have any information specific to this vector, or can suggest a vector (from personal experience) in the collection for ubiquitous expression of a gene::GFP fusion (preferably with Not, XMA and KPN1 sites for cloning), please let me know.

Edit: Sorry if something like this is covered in previous posts, for some reason the search feature doesn’t seem to be working on this browser.

Thanks,
Joe

2

Have you tried the documentation for the Fire vectors, available at Addgene? The Decoy sequence you ask about is described in the 1995 kit documentation, and the plasmid you are using is described in the 2005 kit documentation (as L5494).

Sadly the search function has been broken for a while, but you can always do a Google search including the term “site=forums.wormbase.org”.

3

Thank you so much. That’s exactly the information I was looking for, I don’t know how I couldn’t find it before. Unfortunately, it doesn’t seem to offer a clue as to what I’m doing wrong, however, I feel much more comfortable now that I understand the vector better. I may just need to inject a whole lot more worms.