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:	DIAZ CARRASCO, ASUNCION, CENTRO DE INVEST. BIOL., VELASQUEZ, 144, E-28006 MADRID, ES, LOPEZ GARCIA, PALOMA, CENTRO DE INVES. BIOLOGICAS, VELAZQUEZ, 144, E-28006 MADRID, ES, LACKS, SANFORD, A., UPTON, NY 11973, US, ESPINOSA PADRON, MANUEL, CENTRO DE INVEST. BIOL., VELASQUEZ, 144, E-28006 MADRID, ES, PONS SALVADOR, M ELENA, CENTRO DE INVES.BIOL.DEL, ELASQUES, 144, 28006 MADRID, ES
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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Beschreibung
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.