Molecular Cloning of Bovine Thyroglobulin Complementary DNA. Characterization of 2500-Base-Pair and 1900-Base-Pair Fragments
| Autor: | Etienne Pays, Huguette Brocas, Frank Gannon, Daniel Christophe, Guy De Martynoff, Gilbert Vassart |
|---|---|
| Rok vydání: | 1980 |
| Předmět: |
endocrine system
Transcription Genetic Base pair medicine.medical_treatment DNA Recombinant Deoxyribonuclease HindIII Biology Molecular cloning Thyroglobulin Biochemistry law.invention chemistry.chemical_compound Plasmid law Complementary DNA Escherichia coli medicine Animals RNA Messenger Cloning Molecular Base Composition Nucleic Acid Hybridization DNA Restriction Enzymes Molecular biology Restriction enzyme chemistry Polyribosomes Protein Biosynthesis Recombinant DNA Cattle DNA Plasmids |
| Zdroj: | European Journal of Biochemistry. 111:419-423 |
| ISSN: | 1432-1033 0014-2956 |
| Popis: | Double-stranded thyroglobulin complementary DNA (cDNA) was synthesized from purified 33-S bovine thyroglobulin mRNA. This synthetic structural gene has previously been shown to contain three sites for the restriction endonuclease HindIII, yielding two internal fragments of 1900 and 2500 base pairs respectively. Recombinant molecules were prepared by ligating the HindIII-restricted cDNA to the plasmid pBR322 which had been linearized by the same enzyme. When Escherichia coli was transformed with this mixture, it yielded two kinds of colonies each harboring recombinant plasmids containing one of the two cDNA fragments. Both recombinant molecules hybridized specifically to translatable thyroglobulin mRNA. Sequence homology between the two cloned DNAs could not be detected by cross-hybridization experiments; this argues against the existence of internal structural repetition in thyroglobulin subunits. Together, the two cloned DNA fragments represent 55% of the 8000-base-pair double-stranded thyroglobulin DNA. |
| Databáze: | OpenAIRE |
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