Purification of major abundance class of poly(A+)-RNA from rat ventral prostate

Per Erik Mansson, Alan B. Silverberg, Sandra H. Gipson, Stephen E. Harris

Producción científica: Articlerevisión exhaustiva

3 Citas (Scopus)


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 32P-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°C. HAP purified double-stranded material was 92% resistant to S1 nuclease. The DNA-DNA reannealing profile of double stranded 32P-cDNA enriched for the 500 NT band gave a Cot1/2of ~7 × 10−4moles × sec × 1−1 indicating a complexity for this enriched synthetic gene of 500–600 nucleotide pairs (NTP).

Idioma originalEnglish (US)
Páginas (desde-hasta)229-241
Número de páginas13
PublicaciónMolecular and Cellular Endocrinology
EstadoPublished - 1980
Publicado de forma externa

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology


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