Purification of mRNA coding for rat-liver fructose-1,6-bisphosphatase by polysome immunoabsorption.
The mRNA coding for rat liver fructose-1,6-bisphosphatase, which represents approx. 0.46% of total hepatic mRNA, has been purified to near homogeneity. Polysomes from rat liver were allowed to react with antibodies to rabbit anti-fructose-1,6-bisphosphatase purified by affinity chromatography. The complex was immobilized on a protein A-Sepharose column. After the removal of unabsorbed polysomes, the specific mRNA was eluted and chromatographed on an oligo(dT)-cellulose column. This method gave a 183-fold enrichment of the fructose-1,6-bisphosphatase mRNA to greater than 80% homogeneity as determined by electrophoreses of immunoprecipitated in vitro translation products on polyacrylamide slab gels in the presence of sodium dodecyl sulphate.
D000818:Animals; D002474:Cell-Free System; D006597:Fructose-Bisphosphatase / Q000235:genetics* / Q000276:immunology; D007158:Immunologic Techniques; D008099:Liver / Q000201:enzymology / Q000502:physiology; D011132:Polyribosomes / Q000276:immunology; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D051381:Rats
D012333:RNA, Messenger; D006597:Fructose-Bisphosphatase