Cloning in Escherichia coli of the structural gene of prorennin, the precursor of calf milk-clotting enzyme renin.

Double-stranded cDNA was prepared from prorennin-specific mRNA by sequential actions of reverse transcriptase, DNA polymerase and S1 nuclease, and inserted into the Sa/I site of pBR322 by the poly(dG)-(dC) annealing method. Transformation of Escherichia coli C600 r- m- by the hybrid plasmid yielded transformants containing prorennin cDNA. The presence of the cDNA sequence in these clones was confirmed by both colony hybridization and hybrid-arrested translation of the mRNA in vitro. The largest size of the cloned cDNA was 1,020 bp.