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P Commer

Publications and source records attributed to P Commer.

3 recordsLinked to original sources
PubMed1979

alpha 1-Fetoprotein mRNA of rat yolk sac and hepatoma.

Rat alpha 1-fetoprotein mRNA was isolated and purified to apparent homogeneity by means of immunoadsorption and oligo (dT) cellulose affinity chromatography. Purified AFP mRNA migrated as a 21S peak in 2.5% SDS-polyacrylamide gels. The translation product of this mRNA in micrococcal nuclease treated reticulocyte lysate was identified as AFP by specific immunoprecipitation, SDS-gel electrophoresis and tryptic digestion analysis. DNA complimentary to AFP mRNA was synthesized with avian meyloblastosis virus RNA-dependent DNA polymerase. This AFP cDNA was used as a probe to quantitate AFP mRNA in the developing rat liver and to compare the complexity and diversity of AFP mRNA derived from the normal rat liver and Morris hepatoma 7777. We found that the amount of functional AFP mRNA is decreasing during liver development. There is very little, if any, AFP mRNA in the adult rat liver. A high degree of homology between the AFP mRNA sequences of yolk sac and hepatoma was also found.

Animals↗
PubMed1979

Isolation of rat alpha1-fetoprotein messenger RNA from Morris hepatoma 7777.

A double-antibody procedure has been developed for the isolation of alpha1-fetoprotein (AFP)-synthesizing polysomes from Morris hepatoma 7777. The polyadenylic acid-containing RNA, subsequently purified by differential sedimentation on sucrose gradient and oligodeoxythymidylic acid-cellulose chromatography, migrates as a single 21S component in polyacrylamide gel electrophoresis; in a cell-free translation system, it yields a peptide product immunoprecipitable by anti-rat AFP antiserum, but not by anti-rat albumin, and which migrates slightly faster than serum AFP on sodium dodecyl sulfate-urea-polyacrylamide gels. This messenger RNA fraction was used for the synthesis of a radioactive complementary DNA. In hybridization assays, the complementary DNA reassociated with its purified template at a Cr0t1/2 [product of RNA concentration (mol of nucleotides per liter) X half-time (sec)] of 1.5 X 10(-2). By constitute 3,2, and less than 0.01% of total polyadenylic acid-containing polysomal RNA of Morris hepatoma 7777, 10-day-old-rat liver, and adult rat liver, respectively. The high specificity of the polysome immunoprecipitation system, the electrophoretic homogeneity of the isolated messenger RNA fraction, its selective translation into AFP, and the specificity of the hybridization probe indicate that the procedure described yields a highly purified rat AJP messenger RNA.

Animals↗