PMID: 6989688Mar 1, 1980Paper

Isolation and characterization of ovine prolactin and growth-hormone messenger RNAs

Molecular and Cellular Endocrinology
M A Shupnik, J Gorski

Abstract

Methods are described in this paper for obtaining and characterizing highly enriched preparations of ovine prolactin (PRL) and growth-hormone (GH) messenger RNAs. Purification steps include phenol extraction, oligo-(dT)-cellulose chromatography, sucrose-gradient ultracentrifugation, and polyacrylamide-gel electrophoresis. This sequence of procedures results in messenger preparations that are about 92% pure for prolactin mRNA and 78% pure for the growth-hormone mRNA species. Thus, these two closely related mRNAs can be isolated from the same tissue source at a purity adequate for cloning and nucleic acid hybridization experiments. Translation experiments with the cap analogue 7-methylguanosine, and end-labeling of the nucleic acid before and after beta-oxidation indicate that both messages possess blocked 5'-termini, and that these are part of previously described cap structures. Polyadenosine tracts of 30-160 residues were found at the 3'-ends of both purified species. Finally, sizing experiments suggest both mRNAs contain approx. 30% more bases than accounted for by the coding regions and the poly(A)-tracts. Their physical characteristics thus agree with those of most eucaryotic messages to date.

References

Dec 1, 1977·Nucleic Acids Research·A EfstratiadisF C Kafatos
Feb 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A M Maxam, W Gilbert
Jun 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·R J TushinskiF C Bancroft
Jun 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·V R LingappaG Blobel
Mar 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·G A EvansM G Rosenfeld
Aug 1, 1976·European Journal of Biochemistry·H R Pelham, R J Jackson
Aug 5, 1974·Biochemical and Biophysical Research Communications·H H KazazianT C Cheng
Jun 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·H Aviv, P Leder
Jun 4, 1974·Biochemistry·V GlisinC Byus

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