Screening of Proteins Interacting with Cocaine-and Amphetamine-Regulated Transcript Peptide
|School||Second Military Medical University|
|Keywords||cDNA library CART Bacterial two -hybrid system Protein - protein interactions|
[Abstract ] Objective: To find the protein can Kayin An amphetamine regulated transcript peptide (CART) receptor or its interaction . Methods: Rat brain cDNA library to CART 41-102 as bait , using bacterial two-hybrid screening CART interacting protein . The obtained positive results for DNA sequencing , further analyze the positive results based on bioinformatics methods . Results : The successful construction of a rat brain cDNA library and bait plasmid pBT -CART . Library capacity of 3.37 × 10 6 sup>, recombination rate of 98.5% . Screening approximately 5.01 × 10 6 sup> co-transformed clones, get positive clones 125 , confirmed 93 positive clones , send sequencing 93 . Analyzed by bioinformatics methods stathmin 6 may be interacting protein and CART , and get a new and unknown protein may interact and CART DNA sequences, which are 22 known membrane proteins or receptors very similar short fragment. Conclusion : The bacterial two - hybrid system is a simple and effective method of interaction between a protein identification . CART may interact with stathmin six known protein and 22 unknown protein interactions , these unknown protein with 22 membrane protein or receptor short similarity conservative structure .