PMID: 6171425Nov 1, 1981Paper

Cloning of four DNA fragments complementary to human thyroglobulin messenger RNA

European Journal of Biochemistry
J L Bergé-LefrancS Lissitzky

Abstract

Human thyroglobulin mRNA was isolated from Graves' goitres by size selection of total poly(A)-rich RNA in a sucrose gradient. It sedimented at 33 S, as in other mammalian species, and showed a single component of approximately 8500 bases by gel electrophoresis. cDNA was synthesized from the 33-S RNA by using reverse transcriptase in the presence of human placenta ribonuclease inhibitor and in conditions allowing the formation of long transcripts. The latter was made double-stranded using reverse transcriptase and blunt-ended with nuclease S1. After tailing with dCTP and terminal transferase, the double-stranded cDNA was annealed to pBR322 DNA that had been cleaved at the endonuclease PstI site and tailed with dGTP. The resulting plasmids were used to transform Escherichia coli C600 cells and four cloned recombinants were selected. Each plasmid DNA was shown to contain a sequence complementary to human thyroglobulin mRNA by hybridization with a labeled 33-S mRNA, visualization of cDNA . mRNA hybrids by electron microscopy and filter hybridization selection of mRNA directing the synthesis of immunologically related thyroglobulin peptides in the reticulocyte lysate. The four inserted DNA sequences were 1400 - 1800 base pairs long,...Continue Reading

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Citations

Jan 1, 1985·Human Genetics·J L Bergé-LefrancS Lissitzky
Feb 1, 1989·Biochimie·Y MalthièryS Lissitzky
May 1, 1985·Molecular and Cellular Endocrinology·G VassartH Targovnik
Jan 1, 1991·Thyroid : Official Journal of the American Thyroid Association·M NoelM S Moukhtar
Jan 1, 1991·Thyroid : Official Journal of the American Thyroid Association·H M TargovnikG Medeiros-Neto
May 20, 1998·Thyroid : Official Journal of the American Thyroid Association·H M TargovnikG Medeiros-Neto
Apr 25, 1983·Nucleic Acids Research·G J van OmmenJ J de Vijlder
Apr 25, 1984·Nucleic Acids Research·V E AvvedimentoR Di Lauro
Jun 1, 1984·European Journal of Biochemistry·H M TargovnikG Vassart
Jun 15, 1987·European Journal of Biochemistry·Y Malthiéry, S Lissitzky
Feb 1, 1984·Journal of Endocrinological Investigation·S Lissitzky
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
Jan 1, 1990·Autoimmunity·J Furmaniak, B R Smith
Jun 1, 1993·Journal of Endocrinological Investigation·H M TargovnikG A Medeiros-Neto
Mar 11, 2021·Molecular and Cellular Endocrinology·Cintia E CitterioHéctor M Targovnik
Apr 14, 1982·Biochemical and Biophysical Research Communications·D ChristopheG Vassart

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