Screening Of Proteins Interacting With Cocaine-and Amphetamine-Regulated Transcript Peptide

The objective was to looking for CART receptors or proteins interacted to cocaine-and amphetamine — regulated transcript peptide(CART). A cDNA library from rat brain was constructed. Then the BacterioMatch two-hybrid system was used to screening for proteins interacted with CART, using CART41.102 as bait, and rat brain cDNA library as the pool from which the interactors were draw. The cDNA library and bait pBT- CART41.102 was constructed successfully. The library harvested 3. 37×10~6 individuals, and the reconstruction rate was 98.5%. 5.01xl06 cotransformed clones were screened, 93 clones were conformtively identified to interacting with CART. All were DNA sequenced. By bioinformation analysis, 6 known proteins interacted to CART were identified. Other unknown interaction proteins that had short exactly similiar motif to 22 membraneproteins or receptors were also identified. CART might interact to proteins about energy balance, cell proliferation and enzymes related to protein processing and degrading .