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The expression of winter flounder liver-type antifreeze protein (wflAFP) genes is tissue-specific and under seasonal and hormonal regulation. The only intron of the major wflAFP gene was demonstrated to be a liver-specific enhancer in both mammalian cell lines and flounder hepatocytes. Element B, the core enhancer sequence, was shown to interact specifically with a liver-enriched transcription factor, CCAAT/enhancer-binding protein [alpha] (C/EBP[alpha]), as well as a presumptive antifreeze enhancer-binding protein (AEP). In this study, the identity of the rat AEP ortholog was revealed via its DNA–protein interaction with element B. It is a helicase-domain-containing protein, 988 amino acids in length, and is homologous to mouse Smubp-2, hamster Rip-1 and human Smubp-2. The specific binding between element B and AEP was confirmed by South-Western analysis and gel retardation assays. Residues in element B important to this interaction were identified by methylation interference assays. Mutation on one of the residues disrupted the binding between element B and AEP and its enhancer activity was significantly reduced, suggesting that AEP is essential for the transactivation of the wflAFP gene intron. The rat AEP is ubiquitously expressed in various tissues, and the flounder homolog is present as shown by genomic Southern analysis. The potential role of AEP in regulating the flounder AFP gene expression is discussed.
Keywords: antifreeze protein (AFP); DNA-binding protein; antifreeze enhancer-binding protein (AEP); helicase-domain-containing protein; transcriptional regulation.
Abbreviations: AFP, antifreeze protein; wflAFP, winter flounder liver-type AFP; wfsAFP, winter flounder skin-type AFP; C/EBP[alpha], CCAAT/enhancer-binding protein alpha; AEP, antifreeze enhancer-binding protein; GH, growth hormone; AP-1, activator protein-1; MBP, maltose-binding protein; HNF3, hepatocyte nuclear factor 3; CAT, chloramphenicol acetyltransferase
Originally posted by dacruz
(Isn´t that against the law to modify a DNA?)
Originally posted by klain
ok so your saying that if some weird crazy nut probably sum1 like me could get away with creating a half man half goat hybrid i knw some animals have some of ours like chimaras or what ever their called