In recent years, the general public has recognized the dangers of a high fat diet and
are demanding meat with lower fat content. This demand has stimulated research in the
growth and regulation of adipocytes. However, despite much effort, no adipocyte-specific
plasma membrane markers from any species are available as an aid to accurately distinguish
adipocytes from non-adipocytes. One potential candidate for such a marker in
porcine adipocytes has been identified by Killefer and Hu (1990b). Characterization of the
cDNA for this protein, designated porcine adipocyte membrane protein (PAMP), is presented
here. Sequence for the 910 by clone is 80% similar to an internal region of a rat
prostaglandin F[subscript 2��] receptor regulator protein (FPRP) described by Orlickey (1996). Western
blot analysis suggests that the pig protein is a homotetramer held together with disulfide
bonds which form very close to the transmembrane region making the tetramer
extremely difficult to reduce to monomeric units. Oligonucleotide primers were designed
to amplify a genomic fragment by the polymerase chain reaction (PCR) and for a reverse
transcriptase PCR (RT-PCR) assay to study the expression of the mRNA. A 2114 bp
genomic clone revealed one intron in the coding region. A serum-free primary cell culture
system was used to study the expression of the mRNA. Although message was detected
every day over a ten day period, it appeared to peak between 6 to 8 days after plating.
The PAMP protein is clearly of the same family as the rat FPRP but its size and conformation
are quite different so it is not clear what function it performs in porcine adipocytes.
Further experiments should focus on attaining full length cDNA's, confirming the molecular
conformation of the protein, and assessing its function in a serum-free primary cell
culture system. / Graduation date: 1997
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/34266 |
| Date | 02 May 1997 |
| Creators | Vergin, Kevin L. |
| Contributors | Hu, Ching Yuan |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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