FIELD: molecular biology, biotechnology. SUBSTANCE: plasmid pAE12 is constructed on the basis of pKK233-2 by digestion with restrictases NeoI and HindIII and ligation to DNA-fragment encoding an alpha-amidating enzyme. Plasmid has hybrid trp-lac. Strain Escherichia coli JM 105 is transformed with plasmid pAE12. Expression vector for murine cells C127 is constructed on the basis of pdMMTNeo. Obtained plasmid pdBPV-MMTNeo αAEA75 has a promoter MMT, DNA-sequence encoding an enzyme and a gene determining resistance to neomycin. Murine cellular line C127 is transformed with indicated plasmid and a line - a producer of an alpha-amidating enzyme is obtained. Invention can be used for preparing an alpha-amidating enzyme. EFFECT: high yield and purity of an enzyme. 5 cl, 4 dwg, 2 ex
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1989 |
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