Purification and characterization of procollagen mRNA from a heavy polysomal fraction.
A method is described for the purification of intact procollagen mRNA from heavy polysomal fractions isolated from 10 day-old chick embryos. Procollagen mRNA activity was characterized by gel electrophoresis under denaturing conditions and by its activity in in vitro cell-free translation systems derived from wheat germ and rabbit reticulocytes. 7-Methyl-guanosine monophosphate inhibited the translation of procollagen mRNA and globin mRNA to the same extent. Collagenase-sensitive polypeptides synthesized displayed the same electrophoretical mobilities as marker procollagen chains. The purity of the mRNA was 78% on the basis of ROt plot analysis of hybridization with its corresponding cDNA, using globin cDNA as the reference for 100%.
D000818:Animals; D002458:Cell Fractionation; D002642:Chick Embryo; D004270:DNA, Circular / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D011132:Polyribosomes / Q000378:metabolism*; D011347:Procollagen / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification*; D011817:Rabbits
D004270:DNA, Circular; D011347:Procollagen; D012333:RNA, Messenger