Purification of pro rennin messenger rna and its translation in vitro
Uchiyama, H.; Uozumi, T.; Beppu, T.; Arima, K.
Agricultural and Biological Chemistry 44(6): 1373-1382
ISSN/ISBN: 0002-1369 Accession: 006233417
Prorennin-specific mRNA was 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, 4th 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, on addition of this mRNA, was identified as authentic prorennin by gel electrophoresis. The MW of this mRNA was about 3.5 .times. 105 as determined by gel electrophoresis. The synthesis of prorennin is directed by this mRNA 1020 nucleotides in length and requires the full coding capacity of the molecule. [Prorennin is a precursor of rennin (EC 22.214.171.124), a milk clotting enzyme used in cheese making.].