Translation in a cell-free system of mRNA from term placenta extracted by guanidine hydrochloride.
Human term placenta RNA and polyadenylated mRNA were prepared using guanidine HCl and oligo (dT)-cellulose affinity chromatography. Both fractions translated in a wheat germ cell-free system showed, under optimal condition of K+ and Mg++ ions and spermidine, about 9 times activity for RNA and 15-25 times for poly(A+) mRNA greater than the control. Homogenization of the tissue at high speed compared to that at low speed improved 4-fold activity. Analysis of tritiated products by SDS-Polyacrylamide gel electrophoresis and detected by fluorography showed more than ten different intensity bands ranging between 12 and 66 kD. According to the results obtained, guanidine HCl is an advantageous procedure for the extraction of RNA from this nuclease-rich tissue compared with that obtained with phenol extraction, in both activity and in larger translation products.
D000818:Animals; D002474:Cell-Free System; D005260:Female; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D008099:Liver / Q000032:analysis; D010920:Placenta / Q000032:analysis*; D011247:Pregnancy; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats
D006146:Guanidines; D012333:RNA, Messenger; D019791:Guanidine