Cloning and sequence analysis of the human obesity-related neuropeptide Y(NPY)Y2 receptor gene

Abstract

Objective The present paper aims at cloning and identifying human neuropeptide Y receptor Y2 ( NPY2R), one of the human obesity - related genes. Methods The cDNA of NPY2R gene was amplified from RNA of the human fat by reverse transcription polymerase chain reaction ( RT - PCR) . The PCR products were recombined with pET28a + vector, positive clones were selected and DNA sequence analysis was conducted for the identification, followed by sequence analysis and homology comparison with that reported in Genebank after sequencing. Results A DNA fragment of about 1 100 bp was obtained by RT- PCR, the cDNA sequence of the recombinant plasmid of pET28a + with the fragment was analyzed, and the result showed that the cloned DNA fragment was just human NPY2R cDNA. The cloned gene was encoded 381 amino acids with a molecular weight of 42 kD. The nucleotide sequence homology of the cloned NPY2R was 100% in comparison with that reported in Genebank. Conclusion The sequence of NPY2R cDNA that we have successfully cloned is the same as that of NPY2R in Genebank, which lays the foundation for further studies of gene expression of NPY2R and the interrelationship of NPY2R and the genesis, development and transformation of obesity by means of molecular biology. Key words: Obesity; Neuropeptide Y (NPY) : Y2 receptor(NPY2R) ; Gene cloning. DNA sequence analysis