A rapid procedure for purification of EcoRI endonuclease

János Sümegi; Danielle Breedveld; paul Hossenlopp; Pierre Chambon

doi:10.1016/0006-291X(77)91670-9

A rapid procedure for purification of EcoRI endonuclease

János Sümegi, Danielle Breedveld, paul Hossenlopp, Pierre Chambon

Pathology & Microbiology

Research output: Contribution to journal › Article › peer-review

43 Scopus citations

Abstract

A convenient and rapid procedure has been developed to purify restriction endonuclease Eco RI. The method involves sonication of cells at low ionic strength, precipitation of the endonuclease with Polymin P (a polyethyleneimine), elution of the enzyme from the Polymin P precipitate, ammonium sulfate precipitation and chromatography on phosphocellulose. The purified restriction endonuclease is free of exonuclease and other endonucleases.

Original language	English (US)
Pages (from-to)	78-85
Number of pages	8
Journal	Biochemical and Biophysical Research Communications
Volume	76
Issue number	1
DOIs	https://doi.org/10.1016/0006-291X(77)91670-9
State	Published - May 9 1977

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "A rapid procedure for purification of EcoRI endonuclease",

abstract = "A convenient and rapid procedure has been developed to purify restriction endonuclease Eco RI. The method involves sonication of cells at low ionic strength, precipitation of the endonuclease with Polymin P (a polyethyleneimine), elution of the enzyme from the Polymin P precipitate, ammonium sulfate precipitation and chromatography on phosphocellulose. The purified restriction endonuclease is free of exonuclease and other endonucleases.",

author = "J{\'a}nos S{\"u}megi and Danielle Breedveld and paul Hossenlopp and Pierre Chambon",

note = "Funding Information: We thank Dr. J.L. Mandel for helpful discussion and J.P. LePennec for a giftof T4 DNA-ligase. One of us (J. Stimegi) is an UNESCO/UNDP fellow on leave from B.R.C. Institute of Biochemistry, Szeged, Hungary. This work was supported by grants from the Institut National de la Sante et de la Recherche Medicale (CRT 76-1462), the Centre National de la Recherche Scientifique (RCP 385) and the Fondation pour la Recherche Medicale Francaise.",

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AU - Sümegi, János

AU - Breedveld, Danielle

AU - Hossenlopp, paul

AU - Chambon, Pierre

N1 - Funding Information: We thank Dr. J.L. Mandel for helpful discussion and J.P. LePennec for a giftof T4 DNA-ligase. One of us (J. Stimegi) is an UNESCO/UNDP fellow on leave from B.R.C. Institute of Biochemistry, Szeged, Hungary. This work was supported by grants from the Institut National de la Sante et de la Recherche Medicale (CRT 76-1462), the Centre National de la Recherche Scientifique (RCP 385) and the Fondation pour la Recherche Medicale Francaise.

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Y1 - 1977/5/9

N2 - A convenient and rapid procedure has been developed to purify restriction endonuclease Eco RI. The method involves sonication of cells at low ionic strength, precipitation of the endonuclease with Polymin P (a polyethyleneimine), elution of the enzyme from the Polymin P precipitate, ammonium sulfate precipitation and chromatography on phosphocellulose. The purified restriction endonuclease is free of exonuclease and other endonucleases.

AB - A convenient and rapid procedure has been developed to purify restriction endonuclease Eco RI. The method involves sonication of cells at low ionic strength, precipitation of the endonuclease with Polymin P (a polyethyleneimine), elution of the enzyme from the Polymin P precipitate, ammonium sulfate precipitation and chromatography on phosphocellulose. The purified restriction endonuclease is free of exonuclease and other endonucleases.

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A rapid procedure for purification of EcoRI endonuclease

Abstract

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