ES2095195A6 - Mejoras introducidas en la patente principal n-9000542 por procedimiento de obtencion y purificacion del fragmento carboxilo-terminal de la dna polimerasa i de streptococcus pneumoniae. - Google Patents

Mejoras introducidas en la patente principal n-9000542 por procedimiento de obtencion y purificacion del fragmento carboxilo-terminal de la dna polimerasa i de streptococcus pneumoniae.

Info

Publication number
ES2095195A6
ES2095195A6 ES09601205A ES9601205A ES2095195A6 ES 2095195 A6 ES2095195 A6 ES 2095195A6 ES 09601205 A ES09601205 A ES 09601205A ES 9601205 A ES9601205 A ES 9601205A ES 2095195 A6 ES2095195 A6 ES 2095195A6
Authority
ES
Spain
Prior art keywords
dna polymerase
purifying
carboxyl
obtaining
terminal fragment
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related
Application number
ES09601205A
Other languages
English (en)
Inventor
Monica Amblar
Asuncion Diaz
Paloma Lopez
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Consejo Superior de Investigaciones Cientificas CSIC
Original Assignee
Consejo Superior de Investigaciones Cientificas CSIC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Consejo Superior de Investigaciones Cientificas CSIC filed Critical Consejo Superior de Investigaciones Cientificas CSIC
Publication of ES2095195A6 publication Critical patent/ES2095195A6/es
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Mejoras introducidas en la patente principal nº 9000542 por Procedimiento de obtención y purificación del fragmento carboxilo-terminal de la DNA polimerasa I de Streptococcus pneumoniae. El procedimiento de obtención y purificación de la DNA polimerasa I D102 de Streptococcus pneumoniae es un méetodo para incrementar la actividad polimerizante del fragmento carboxilo-terminal de la DNA polimerasa I. Este método incluye mutagénesis dirigida del gen po1A, y subclonaje del gen mutado en un vector de expresión de Escherichia coli. Mediante este procedimiento se ha obtenido el plásmido recombinante pMA71. Dicho plásmido codifica un polipéptido de 99 kDa, que posee una actividad polimerizante elevada y que solo retiene menos del 1% de la actividad 5''-3'' exonucleásica de la proteína silvestre. El polipéptido ha sido hiperexpresado y purificado a homogeneidad a partir de E.coli. El polipéptido purificado es soluble y activo, habiéndose obtenido a partir de 2 litros de cultivo 6570 unidades de actividad polimerasa con una actividad específica de 30000 unidades por mg de proteína.
ES09601205A 1990-02-23 1996-05-31 Mejoras introducidas en la patente principal n-9000542 por procedimiento de obtencion y purificacion del fragmento carboxilo-terminal de la dna polimerasa i de streptococcus pneumoniae. Expired - Fee Related ES2095195A6 (es)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES9000542A ES2023538A6 (es) 1990-02-23 1990-02-23 Procedimiento de obtencion y purificacion del fragmento carboxilo-terminal de la dna polimerasa i de streptococcus pneumoniae

Publications (1)

Publication Number Publication Date
ES2095195A6 true ES2095195A6 (es) 1997-02-01

Family

ID=8266083

Family Applications (2)

Application Number Title Priority Date Filing Date
ES9000542A Expired - Fee Related ES2023538A6 (es) 1990-02-23 1990-02-23 Procedimiento de obtencion y purificacion del fragmento carboxilo-terminal de la dna polimerasa i de streptococcus pneumoniae
ES09601205A Expired - Fee Related ES2095195A6 (es) 1990-02-23 1996-05-31 Mejoras introducidas en la patente principal n-9000542 por procedimiento de obtencion y purificacion del fragmento carboxilo-terminal de la dna polimerasa i de streptococcus pneumoniae.

Family Applications Before (1)

Application Number Title Priority Date Filing Date
ES9000542A Expired - Fee Related ES2023538A6 (es) 1990-02-23 1990-02-23 Procedimiento de obtencion y purificacion del fragmento carboxilo-terminal de la dna polimerasa i de streptococcus pneumoniae

Country Status (5)

Country Link
EP (1) EP0470230B1 (es)
AT (1) ATE154074T1 (es)
DE (1) DE69126379T2 (es)
ES (2) ES2023538A6 (es)
WO (1) WO1991013153A1 (es)

Also Published As

Publication number Publication date
ATE154074T1 (de) 1997-06-15
DE69126379D1 (de) 1997-07-10
ES2023538A6 (es) 1992-01-16
WO1991013153A1 (es) 1991-09-05
DE69126379T2 (de) 1997-11-06
EP0470230B1 (en) 1997-06-04
EP0470230A1 (en) 1992-02-12

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Effective date: 20110225