Cloning And Sequening Of Porcine Adipocyte Membrane Protein Gene CDNA

The product of PAMP gene is a 64 KD membrane protein which was identified by Killefer and Hu in 1990.They found the differential expression of the 64KD protein having different genetic background in adipocyte plasma membrane.A monoclonal antibody(LA-l)was used to confirm the presence of the 64KD protein in genetically lean adipocyte plasma membranes but absence in genetically obese adipocyte plasma membranes.The result implies that the protein may be involed in adipocyte regulation such that the lack of the protein may lead to obesity.The total RNA was extracted from porcine backfat adipose tissue.A primer was designed according to the 3'end cDNA sequence of PAMP gene (Hu,1997).Based on the method of RACE,530bp fragment of PAMP gene 5'end cDNA was amplified and cloned by T/A vector. There is no homologous sequence between PAMP gene cDNA and FPRP gene cDNA. It is identical with the result of Hu. In their sequence,there are two regions of 45 and 84 nucleotides each with 33% similarity to the FPRP gene in the 3' untranslated region.In the result of this study,there are no similar sequence in the 5' untranslated region.lt further more confirms Hu's prediction:PAMP and FPRP are different proteins which are alternative spliced from the same gene.A primer was designed according to the new sequence of PAMP gene cDNA. Combined with the primer of RACE,It implies that there is no intron in this region of genomic DNA.Cloning and sequencing analysis of PAMP gene cDNA gave us some ideas of the real fuction of PAMP.