Purification of Prorennin mRNA and Its Translation in Vitro

Abstract

Prorennin-specific messenger ribonucleic acid (mRNA) has been purified by a combination of sizing techniques, including Sepharose 2B chromatography and sucrose density gradient centrifugation, and affinity chromatography with poly (U)-Sepharose, from total nucleic acid extracted from dry ice-pulverized, fourth stomach of a calf. This mRNA bound to poly (U)-Sepharose, indicating that it contained a poly (A) sequence. The total translation product in the mRNA-dependent wheat germ system, upon addition of this mRNA, was identified as authentic prorennin by gel electrophoresis. The molecular weight of this mRNA was about 3.5×10⁵ as determined by gel electrophoresis. These results indicate that the synthesis of prorennin is directed by this mRNA 1, 020 nucleotides in length and requires the full coding capacity of the molecule.