I've recently been working on designing chimeras of mouse GPCRs for electrophys. experiments. The "parent" receptors express well, and I believe that my alignment of them was correct. I cannot seem to get any chimeras to work-- I've done some reading, and there is conflicting evidence as to where I should design the "joints" for the chimeras.

Does anyone have any experience with this?? Any help would be greatly appreciated- my dissertation is on standstill until i can figure this out

SRep said:

I've recently been working on designing chimeras of mouse GPCRs for electrophys. experiments. The "parent" receptors express well, and I believe that my alignment of them was correct. I cannot seem to get any chimeras to work-- I've done some reading, and there is conflicting evidence as to where I should design the "joints" for the chimeras. Does anyone have any experience with this?? Any help would be greatly appreciated- my dissertation is on standstill until i can figure this out

Your problem is not very clear.
However, if your problem is the construction of chimera between two GPCRs some reading may be of help:

Dezhong Yin, Shai Gavi, Hsien-yu Wang, and Craig C. Malbon Probing Receptor Structure/Function with Chimeric G-Protein-Coupled Receptors Mol Pharmacol 65:1323-1332, 2004.

In this minirev you'll find a table summarizing the huge work that have been published regarding the studies of receptor structure/function by construction of chimeric GPCRs.
GOOD LUCK.

my actual problem is that ive made several chimeras with joints at several different regions of the GPCRs, and none have worked. I've even tried an insert chimera of TM5. thanks for the article