Heating RNA before cell-free translation is essential for the efficient and reproducible synthesis of several peroxisomal proteins.
Total RNA, extracted with guanidinium thiocyanate from liver of clofibrate-treated rats, was translated in vitro. Heating the RNA at 60 degrees C for 5 min before translation increased the synthesis of three peroxisomal polypeptides 10-100-fold. Preproalbumin synthesis increased 10-fold. Total incorporation of [35S]methionine into proteins merely doubled. Heating is essential for reproducible and adequate translation of mRNAs coding for peroxisomal and some other proteins.
D000818:Animals; D002474:Cell-Free System; D006358:Hot Temperature; D066298:In Vitro Techniques; D008715:Methionine / Q000378:metabolism; D008830:Microbodies / Q000378:metabolism*; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism*; D051381:Rats; D013698:Templates, Genetic
D012313:RNA; D008715:Methionine