Isolationand in vitro translation of leghaemoglobin mRNA from yellow lupin root nodules.
Messenger RNA for leghaemoglobin, extracted from polysomes of yellow lupin root nodules, was purified by chromatography on oligo(dT1-cellulose followed by sucrose-gradient centrifugation. Leghaemoglobin mRNA activity was assayed in a cell-free protein synthesizing system derived from wheat germ. The purified mRNA was sedimented in surcrose gradient in the 9S region which corresponds to a molecular weight of about 225,000. The poly(A) segment was estimated to be 66 nucleotides in length, based on hybridization with [3H]poly(U). Analysis of the translation product using immunoprecipitation and polyacrylamide-gel electrophoresis in denaturing conditions revealed that the product made in vitro was identical with authentic leghaemoglobin.
D002474:Cell-Free System; D007887:Fabaceae / Q000235:genetics*; D006420:Hemeproteins / Q000235:genetics*; D007874:Leghemoglobin / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics; D010946:Plants, Medicinal; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
D006420:Hemeproteins; D007874:Leghemoglobin; D010940:Plant Proteins; D012333:RNA, Messenger