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

AUTOR(ES)

Mansson, Per-Erik

RESUMO

Total poly(A+)-RNA (poly(A+)-RNAtot) was isolated from rat seminal vesicle and its size distribution determined by 70% formamide 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 11S region (poly(A+)11S. 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+)-RNAtot and poly(A+)-RNA11S. Back-hybridization of poly(A+)-RNAtot and poly(A+)-RNA11S 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 sequences present in 30,000-50,000 copies/cell. Invitro translation of poly(A+)-RNA11S resulted in two major polypeptides coded for by the 620 NT long and 540 NT long poly(A+)-RNA respectively.

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