PMID: 9186272May 1, 1997Paper

The revised 8307 base pair coding sequence of human thyroglobulin transiently expressed in eukaryotic cells

European Journal of Endocrinology
S A van de GraafC Ris-Stalpers

Abstract

We developed a transient transfection system for human thyroglobulin (TG) cDNA in both human thyroid cells and in COS-1 cells. Four overlapping TG cDNA fragments were amplified by reverse transcription-PCR from RNA of normal thyroid tissue. The most 5' fragment includes the natural translation initiation site and the sequence encoding the signal peptide (SP). After subcloning, the nucleotide sequence was determined and compared with the published human sequence, resulting in the detection of 30 nucleotide variations. For validation purposes, all variations were screened in 6-12 normal human alleles. Twenty-one were present in all screened alleles and have to be revised in the published nucleotide sequence. Since one variation concerns a triplet insertion, the coding sequence of the mature human thyroglobulin is 8307 nucleotides encoding 2750 amino acids. The TG cDNA constructs were transiently transfected in HTori 3 and COS-1 cells and protein expression was detected using a polyclonal anti-human-TG on fixed cells and after SDS-PAGE. In both cell-lines all four TG protein fragments were expressed. The mannose structures detected on the proteins by lectins and localization after expression in the cells suggest that only the N-te...Continue Reading

Citations

Jun 8, 1999·The Anatomical Record·A K Christensen, C M Bourne
Jun 9, 2001·Thyroid : Official Journal of the American Thyroid Association·J T Dunn, A D Dunn
Dec 17, 2002·Thyroid : Official Journal of the American Thyroid Association·Carina M RivoltaHéctor M Targovnik
Oct 19, 1999·Thyroid : Official Journal of the American Thyroid Association·F M MendiveH M Targovnik
Sep 29, 2005·Thyroid : Official Journal of the American Thyroid Association·Jussara Vono-TonioloPeter Kopp
May 11, 2013·Thyroid : Official Journal of the American Thyroid Association·Yasuhiro MochizukiJun Amano
Dec 7, 2007·Clinica Chimica Acta; International Journal of Clinical Chemistry·Jen-Der Lin
Jul 28, 2006·Clinica Chimica Acta; International Journal of Clinical Chemistry·Carina M Rivolta, Héctor M Targovnik
Jan 23, 2010·Molecular and Cellular Endocrinology·Héctor M TargovnikCarina M Rivolta
Dec 22, 2004·Arquivos brasileiros de endocrinologia e metabologia·Jussara Vono-Toniolo, Peter Kopp

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