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...

Full description

Saved in:
Bibliographic Details
Main Authors 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
LanguageGerman
Published 06.11.1997
Edition6
Subjects
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
Online AccessGet full text

Cover

More Information
Summary: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.
Bibliography:Application Number: DE19916026379T