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Mol Cell Endocrinol ; 19(3): 229-41, 1980 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-6157588

RESUMEN

Poly(A+)--RNA from rat ventral prostate was isolated using oligo(dT)-cellulose chromatography. 45% of the total poly(A+)--RNA was a single peak at 10S as demonstrated by centrifugation in a 5-20% sucrose gradient containing 1% SDS. By using complementary DNA probes, it was shown that the 10S RNA contained the major abundance class of poly(A+)--RNA. Denaturing agarose-gel analysis revealed 2 major bands in the 10S poly(A+)--RNA preparation approx. 600 NT and 500 NT (NT = nucleotides) long, resp. Double-stranded 32 P-DNAs complementary to light side and heavy side of the 10S poly(A+)--RNA peak were synthesized and isolated using reverse transcriptase and hydroxyapatite (HAP) chromatography. Approx. 40% of the first strand of the cDNAs were converted to double-stranded structures with a Tm of 88 degrees C. HAP purified double-stranded material was 92% resistant to S1 nuclease. the DNA--DNA reannealing profile of double stranded 32 P-cDNA enriched for the 500 NT band gave a Cot 1/2 of approximately 7 X 10(-4) moles X sec X 1(-1) indicating a complexity for this enriched synthetic gene of 500-600 nucleotide pairs (NTP).


Asunto(s)
Poli A/aislamiento & purificación , ARN Mensajero/aislamiento & purificación , ARN/aislamiento & purificación , Animales , ADN/análisis , Enzimas de Restricción del ADN/análisis , Masculino , Desnaturalización de Ácido Nucleico , Hibridación de Ácido Nucleico , Próstata/análisis , Ratas
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