Sequence Composition of Rat Nuclear Deoxyribonucleic Acid and High Molecular Weight Nuclear Ribonucleic Acid

David S. Holmes, James Bonner

Resultado de la investigación: Article

50 Citas (Scopus)

Resumen

Rat ascites nuclear DNA has been characterized by its reassociation profile [(Na+) = 0.18 at 62 °] as judged by chromatography on hydroxyapatite, : Single copy DNA (C0t1/2 obed=1.5×103) comprises 65% of the genome. Sequences repeated an average 1800 times (middle repetitive DNA, C0t1/2 obed=1) comprise 19% of the genome while 9% of the genome (highly repetitive DNA) reassociates faster than is measured in these experiments (C0t1/2 obed <2 ×10-2). Middle repetitive and single copy DNA have been isolated and characterized with respect to their reassociation kinetics and melting profiles. Both isolated classes reassociate with the kinetics describing these components in total nuclear DNA. Reassociated single copy DNA has high thermal stability indicative of fidelity in base pairing; reassociated middle repetitive DNA has a lower thermal stability that may be attributed, in part, to base-pair mismatch, and, in part, to the shortness of the duplex regions. Rat ascites heterogeneous nuclear RNA (HnRNA, 5-10 × 106 daltons) has been sheared and hybridized in excess to isolated single copy or middle repetitive DNA [(Na+) = 0.18 at 62°]. HnRNA hybridizes to a minimum of 4.5 % of the single copy and a minimum of 7.8% of the middle repetitive DNA. The Tm of single copy or middle repetitive hybrids is 1-2° lower than that of the reassociated homologous DNA. DNA isolated from single copy or middle repetitive hybrids reassúciates with kinetics similar to the input DNA. Sheared HnRNA has been hybridized to an excess of total nuclear DNA. 38 % of the HnRNA hybridizes with kinetics (C0t1/2 = 2×103) similar to single copy DNA and 12% hybridizes with kinetics (C0t1/2 = 5.6) a little more slowly than the major reassociating component of middle repetitive DNA. The remaining 50% of the HnRNA failed to hybridize.

Idioma originalEnglish
Páginas (desde-hasta)841-848
Número de páginas8
PublicaciónBiochemistry
Volumen13
N.º5
DOI
EstadoPublished - 1 feb 1974

Huella dactilar

Rats
Molecular Weight
Molecular weight
RNA
DNA
Chemical analysis
Kinetics
Genes
Genome
Ascites
Base Pair Mismatch
Thermodynamic stability
Heterogeneous Nuclear RNA
Hot Temperature
Durapatite
Chromatography
Base Pairing
Freezing
Melting

ASJC Scopus subject areas

  • Biochemistry

Citar esto

Holmes, David S. ; Bonner, James. / Sequence Composition of Rat Nuclear Deoxyribonucleic Acid and High Molecular Weight Nuclear Ribonucleic Acid. En: Biochemistry. 1974 ; Vol. 13, N.º 5. pp. 841-848.
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title = "Sequence Composition of Rat Nuclear Deoxyribonucleic Acid and High Molecular Weight Nuclear Ribonucleic Acid",
abstract = "Rat ascites nuclear DNA has been characterized by its reassociation profile [(Na+) = 0.18 at 62 °] as judged by chromatography on hydroxyapatite, : Single copy DNA (C0t1/2 obed=1.5×103) comprises 65{\%} of the genome. Sequences repeated an average 1800 times (middle repetitive DNA, C0t1/2 obed=1) comprise 19{\%} of the genome while 9{\%} of the genome (highly repetitive DNA) reassociates faster than is measured in these experiments (C0t1/2 obed <2 ×10-2). Middle repetitive and single copy DNA have been isolated and characterized with respect to their reassociation kinetics and melting profiles. Both isolated classes reassociate with the kinetics describing these components in total nuclear DNA. Reassociated single copy DNA has high thermal stability indicative of fidelity in base pairing; reassociated middle repetitive DNA has a lower thermal stability that may be attributed, in part, to base-pair mismatch, and, in part, to the shortness of the duplex regions. Rat ascites heterogeneous nuclear RNA (HnRNA, 5-10 × 106 daltons) has been sheared and hybridized in excess to isolated single copy or middle repetitive DNA [(Na+) = 0.18 at 62°]. HnRNA hybridizes to a minimum of 4.5 {\%} of the single copy and a minimum of 7.8{\%} of the middle repetitive DNA. The Tm of single copy or middle repetitive hybrids is 1-2° lower than that of the reassociated homologous DNA. DNA isolated from single copy or middle repetitive hybrids reass{\'u}ciates with kinetics similar to the input DNA. Sheared HnRNA has been hybridized to an excess of total nuclear DNA. 38 {\%} of the HnRNA hybridizes with kinetics (C0t1/2 = 2×103) similar to single copy DNA and 12{\%} hybridizes with kinetics (C0t1/2 = 5.6) a little more slowly than the major reassociating component of middle repetitive DNA. The remaining 50{\%} of the HnRNA failed to hybridize.",
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Sequence Composition of Rat Nuclear Deoxyribonucleic Acid and High Molecular Weight Nuclear Ribonucleic Acid. / Holmes, David S.; Bonner, James.

En: Biochemistry, Vol. 13, N.º 5, 01.02.1974, p. 841-848.

Resultado de la investigación: Article

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