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alpha 1-Fetoprotein mRNA of rat yolk sac and hepatoma
Abstract:
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.
Insights
Researchers isolated rat alpha 1-fetoprotein mRNA to study its levels during liver development and in hepatoma. They found functional alpha 1-fetoprotein mRNA decreases with development, with high homology between yolk sac and hepatoma sequences.
Area of Science:
- Molecular Biology
- Biochemistry
- Developmental Biology
Background:
- Alpha 1-fetoprotein (AFP) is a major fetal protein in rodents.
- Understanding AFP gene expression is crucial for studying liver development and cancer.
Purpose of the Study:
- To isolate and characterize rat alpha 1-fetoprotein mRNA (AFP mRNA).
- To quantify AFP mRNA levels during rat liver development.
- To compare AFP mRNA complexity in normal liver and Morris hepatoma 7777.
Main Methods:
- Immunoadsorption and oligo (dT) cellulose affinity chromatography for mRNA purification.
- SDS-polyacrylamide gel electrophoresis and immunoprecipitation for AFP identification.
- Synthesis of complementary DNA (cDNA) for AFP mRNA and its use as a molecular probe.
Main Results:
- Purified AFP mRNA was identified as a 21S RNA species.
- Functional AFP mRNA levels decrease significantly during rat liver development.
- High sequence homology was observed between AFP mRNA from yolk sac and hepatoma.
Conclusions:
- AFP mRNA is abundant during fetal development and declines sharply postnatally.
- Morris hepatoma 7777 exhibits significant AFP mRNA expression, similar to fetal tissues.
- This study provides insights into AFP gene regulation during liver differentiation and oncogenesis.