Immunological detection of cDNA clones encoding the uncoupling protein of brown adipose tissue: evidence for an antigenic determinant within the C-terminal eleven amino acids.
Poly(A)+RNA was isolated from brown adipose tissue of cold acclimated rats and a fraction enriched for uncoupling protein mRNA was used to generate a cDNA library in pBR 322. Immunological screening of 1,500 colonies with an affinity-purified antiserum against the uncoupling protein yielded five positive clones, pUCPrat1-5. Clone pUCPrat2 encoded the C-terminal 54 amino acids of rat uncoupling protein and exhibited 90% amino acid homology with the hamster protein. Clones pUCPrat3-5 encoded only the C-terminal 11 amino acids suggesting that an antigenic determinant lies within this sequence.
R G
RG
Ridley
H V
HV
Patel
C L
CL
Parfett
K A
KA
Olynyk
S
S
Reichling
K B
KB
Freeman
D000273:Adipose Tissue / Q000032:analysis* / Q000648:ultrastructure; D002001:Adipose Tissue, Brown / Q000032:analysis*; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003080:Cold Temperature; D003101:Collodion; D006224:Cricetinae; D004247:DNA; D000939:Epitopes / Q000032:analysis; D006824:Hybridization, Genetic; D011233:Precipitin Tests; D012313:RNA / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D012689:Sequence Homology, Nucleic Acid
D000939:Epitopes; D012313:RNA; D003101:Collodion; D004247:DNA