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.
D000818:Animals; D002417:Cattle; D012085:Chymosin / Q000096:biosynthesis* / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism; D004792:Enzyme Precursors / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D015719:Single-Strand Specific DNA and RNA Endonucleases
D004792:Enzyme Precursors; D012333:RNA, Messenger; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase; D004720:Endonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases; C032443:prorennin; D012085:Chymosin