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Partial isolation and identification of hepatic stimulator substance mRNA extracted from human fetal liver
Abstract:
AIM:To partially isolate and identify hepatic stimulator substance mRNA from human fetal liver tissues.METHODS:The poly (A)mRNA was extracted from human fetal liver tissues of 4-5 month gestation, fractionated by size on sucrose gradient centrifugation, translated into protein from each fraction in vitro and then its products were tested for HSS activity.RESULTS:Twenty-two 500 total RNA was obtained from human fetal liver tissues and pooled.mRNA of 420 was yielded,processed by oligo(dT)-cellulose column chromatography, then was size-fractionated by ultracentrifution on a continuous sucrose density gradient (5%-25%), and separated into 18 fractions.Translated products of mRNA in fraction 8 and 9 could produce a two-fold increase in the incorporation of( 3)H-TdR into DNA of SMMC-7721 hepatoma cells and in a heat resistant and organ-specific way.CONCLUSION:The partially purified HSS mRNA was obtained and this would facilitate the cloning of HSS using expression vectors.
Insights
Researchers isolated hepatic stimulator substance (HSS) mRNA from human fetal liver. This partially purified mRNA can be used to clone HSS, aiding further research into liver cell growth regulation.
Area of Science:
- Molecular Biology
- Hepatology
- Gene Expression
Background:
- Hepatic Stimulator Substance (HSS) plays a crucial role in liver cell proliferation.
- Identifying and isolating HSS mRNA is essential for understanding its function and for potential therapeutic applications.
Purpose of the Study:
- To partially isolate and identify messenger RNA (mRNA) encoding Hepatic Stimulator Substance (HSS) from human fetal liver tissues.
- To characterize the biological activity of the isolated mRNA fractions.
Main Methods:
- Extraction of poly (A)mRNA from human fetal liver tissues (4-5 month gestation).
- Size fractionation of mRNA using sucrose gradient ultracentrifugation.
- In vitro translation of mRNA fractions and assessment of Hepatic Stimulator Substance (HSS) activity.
Main Results:
- Partially purified Hepatic Stimulator Substance (HSS) mRNA was obtained from human fetal liver.
- mRNA fractions 8 and 9 demonstrated significant biological activity.
- Translated products from these fractions increased DNA incorporation in hepatoma cells, indicating HSS activity.
Conclusions:
- Partially purified Hepatic Stimulator Substance (HSS) mRNA was successfully isolated.
- This isolation facilitates the cloning of HSS using expression vectors.
- The findings pave the way for further investigation into HSS function and liver regeneration.