| 摘要 |
Messenger RNA of prorennin is extracted from the fourth stomach of calf and the double-stranded prorennin cDNA is prepared therefrom. The prorennin cDNA is cloned into a host microorganism, E. coli C600r~m~, to give clones containing the cDNA insert with variable size as confirmed by the base sequence determination in addition to the colony hybridization and hybrid-arrested translation technique. In vitro recombination of two plasmids obtainable from these clones, pTACR102B6 and pTACR103C2, affords plasmid pCR1001 containing the whole coding sequence for prorennin. On addition of the lac operon of E. coli to pCR1001, plasmid pCR2001 is prepared, which plasmid contains the prorennin cDNA sequence and the promoter requisite for the expression of the prorennin gene in a host cell. Transformation of E. coli C600r~m~ by inserting pCR2001 produces a new E. coli strain, E. coli CR1 fully designated as E. coli CR1(r~m~, thr-, leu-, thi-, pCR2001). This genetically engineered new strain of E. coli, when propagated in aqueous nutrient media, produces prorennin. Thus production by bacteria of prorennin which is a proenzyme and a precursor of the milk coagulating enzyme, calf rennin, has now been accomplished.
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