Cloning in Escherichia coli of the Structural Gene of Prorennin, the Precursor of Calf Milk-Clotting Enzyme Rennin

Abstract
Double-stranded cDNA was prepared from prorennin-specific mRNA by sequential actions of reverse transcriptase, DNA polymerase and SI nuclease, and inserted into the SalI 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.

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