VERFAHREN ZUR HERSTELLUNG UND REINIGUNG DES CARBOXYTERMINALEN FRAGMENTS DER DNA-POLYMERASE I VON STREPTOCOCCUS PNEUMONIAE

VERFAHREN ZUR HERSTELLUNG UND REINIGUNG DES CARBOXYTERMINALEN FRAGMENTS DER DNA-POLYMERASE I VON STREPTOCOCCUS PNEUMONIAE

Disclosed is a method for subcloning a fragment of the gene po1A of S.pneumoniae into an expression vector E.coli. With the disclosed method, it has been possible to obtain the recombinant plasmid pSM10. Said plasmid codes for a polypeptide of 70.6 kDa which has the polymerase activity free of exonu...

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Hauptverfasser: PONS SALVADOR, M ELENA, CENTRO DE INV, SUPERIOR DE INVEST. CIENTIFICAS, ESPINOSA PADRON, MANUEL, CENTRO DE INV, DIAZ CARRASCO, ASUNCION, CENTRO DE INV, DE INVEST. CIENTIFICAS, LACKS, SANFORD, A, LOPEZ GARCIA, PALOMA, CENTRO DE INVES
Format: Patent
Sprache:ger
Schlagworte:
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Zusammenfassung:Disclosed is a method for subcloning a fragment of the gene po1A of S.pneumoniae into an expression vector E.coli. With the disclosed method, it has been possible to obtain the recombinant plasmid pSM10. Said plasmid codes for a polypeptide of 70.6 kDa which has the polymerase activity free of exonuclease activity of the enzyme DNA polymerase-exonuclease of S.pneumoniae. The polypeptide has been hyperexpressed and purified from E.coli. The yield of active enzyme obtained corresponds approximately to 7 % of the cellular proteinic content. 0.73 mg of pure protein has been obtained from 500 ml of culture, with a specific enzyme activity of 13537 units of DNA polimerase per mg of protein.