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 |
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| Authors: | Robert G. Ridley Hasmukh V. Patel Craig L. J. Parfett Kimberly A. Olynyk Susanna Reichling Karl B. Freeman |
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| Affiliation: | (1) Department of Biochemistry, McMaster University, 1200 Main Street West, L8N 3Z5 Hamilton, Ontario, Canada;(2) Cancer Research Laboratory, University of Western Ontario, N6A 5B7 London, Ontario, Canada |
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| Abstract: | 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, pUCPratl–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. |
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| Keywords: | brown adipose tissue cloned cDNA immunoscreening mitochondria uncoupling protein |
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