Characterization of the feline thyroglobulin promoter.
The feline thyroglobulin promoter was identified by a combination of standard polymerase chain reaction (PCR) techniques, using primers designed according to regions of homology in published sequences from other species, then adaptor ligated PCR. A 310 bp fragment of the feline thyroglobulin promoter was generated, including 8 nucleotides of adaptor sequence at the 5' end and, based on the putative transcription start site, 36 nucleotides of the thyroglobulin mRNA (untranslated portion). The homology between the feline promoter sequence (from 193 bp upstream to the putative cap site) and canine, bovine and human sequences was 89%, 81% and 78%, respectively. Transient transfection studies, using reporter constructs in which the feline promoter controlled expression of chloramphenicol acetyl transferase, demonstrated promoter activity in thyroid cells, but no activity in non-thyroid cells. The data presented here demonstrate that the feline thyroglobulin promoter may provide a targeting mechanism for somatic gene therapy of feline thyroid disease.
Blackwood
L
L
Molecular Therapeutics Research Group, Division of Small Animal Clinical Studies, Faculty of Veterinary Medicine, University of Glasgow, Bearsden Road, G61 1QH, Glasgow, UK. L.Blackwood@vet.gla.ac.uk
Onions
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Argyle
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DJ
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002415:Cats / Q000235:genetics*; D002417:Cattle; D002460:Cell Line; D004247:DNA / Q000737:chemistry; D004285:Dogs; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D051381:Rats; D017385:Sequence Homology; D013954:Thyroglobulin / Q000235:genetics*; D013961:Thyroid Gland / Q000378:metabolism; D014162:Transfection
D004247:DNA; D013954:Thyroglobulin