Abstract: Objective  To construct uncoupling proteins（UCPs）chimeras and mutations and provide conditions for study on the functional domains of uncoupling pro
tein family.Methods The gene fragments of UCPs chimera were obtained by routine polymerase chain reaction （PCR）and splicing overlap extension PCR（SOE-PCR）.Then these sequences were respectively cloned into the vector of pCR2.1 and sequenced.The recombinant eukaryotic expression plasmid pcDNA3.1 containing UCPs chimera was constructed by restrictive enzyme ligation.The recombinant plasmid of pcDNAUCP1UCP2,pcDNAUCP2UCP1 and pcDNAUCP3UCP1 were identified by PCR and restrictive enzyme digestion.The mutations of pcDNA-UCP3R167G/pcDNA- UCP3R
E171/172GG were generated by site-directed mutagenesis and then verified by sequencing.Results The identification of target gene by restrictve enzyme and PCR was the same as expectation.The sequence of chimera gene was completely identical with the expectation device.Conclusion
The recombinant eukaryotic expression plasmid containing UCPs chimeras and mutations is constructed successfully.
 

Key words: uncoupling proteins, mitochondria, Ca2+ channel