Open AccessPartial isolation and identification of hepatic stimulator substance mRNA extracted from human fetal liver*
Author(s) -
Yang Xiaoming
Publication year - 1998
Publication title -
world journal of gastroenterology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.427
H-Index - 155
eISSN - 2219-2840
pISSN - 1007-9327
DOI - 10.3748/wjg.v4.i2.100
Subject(s) - messenger rna , fetus , centrifugation , microbiology and biotechnology , rna , in vitro , biology , sucrose , differential centrifugation , hepatic stellate cell , chemistry , chromatography , biochemistry , endocrinology , gene , pregnancy , genetics
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.