Isolation and partial purification of the major abundant class rat seminal vesicle poly(A+-messenger RNA

Total poly(A+)-RNA (poly(A+ )-RNAtot) was isolated from rat seminal vesicle and its size distribution determined By 70+ formaaide 5–25% sucrose density analysis. One major peak was resolved in the 10–13 S region and accounted for ∽35% of the total poly(A+)-RNA applied. Preparative 1% SDS, 5–20% linear sucrose density gradients also resolved a single major peak in the IIS region (poly(A+)lls. Analysis of poly(A+)-RNAtot and poly(A+)-RNA11s under denaturing conditions on 2% agarose gel electrophoresis demonstrated two major components in both poly(A )-RNA populations. Size estimations for these components are 620 and 540 NT respectively. 3H-cDNA was made to both poly(A+)-RNA+tot and poly(A+)-RNAlls- Back-hybridization of poly(A+ )-RNA. and poly(A)-RNAlls to their respective 3H-cDNA revealed a highly aBundant class representing 41% and 85% of the sequences in their respective 3H-cDNA's. The highly abundant class corresponded to 3–5 sequenc.es present in 30,000-50,000 copies/cell. In vitro translation of poly(A )-RNA1ls, resulted in two major polypeptides coded for by the 620 NT long and 540 NT long poly(A+)-RNA respectively.