Preparation and properties of insolubilized restriction endonucleases

Yan-Hwa Wu Lee; Robert W. Blakesley; Leonard A. Smith; Jack G. Chirikjian

doi:10.1093/nar/5.3.679

Preparation and properties of insolubilized restriction endonucleases

Yan-Hwa Wu Lee^*, Robert W. Blakesley, Leonard A. Smith, Jack G. Chirikjian

^*Corresponding author for this work

Department of Biological Science and Technology

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

13 Scopus citations

Abstract

Type II restriction endonucleases Bam HI and Eco RI were covalently coupled to Sepharose. These insolubilized enzymes generated fragment patterns for several viral DNAs identical to those produced by the respective free enzymes. Conditions for optimal activity were similar for both bound and unbound forms of the enzymes. Insolubilization improved thermal stability of Bam HI and Eco RI. The bound enzyme can be recovered from reaction mixtures and reused several times. Upon storage at 4°C, coupled endonucleases remained stable for several months.

Original language	English
Pages (from-to)	679-689
Number of pages	11
Journal	Nucleic acids research
Volume	5
Issue number	3
DOIs	https://doi.org/10.1093/nar/5.3.679
State	Published - 1 Mar 1978

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AU - Wu Lee, Yan-Hwa

AU - Blakesley, Robert W.

AU - Smith, Leonard A.

AU - Chirikjian, Jack G.

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N2 - Type II restriction endonucleases Bam HI and Eco RI were covalently coupled to Sepharose. These insolubilized enzymes generated fragment patterns for several viral DNAs identical to those produced by the respective free enzymes. Conditions for optimal activity were similar for both bound and unbound forms of the enzymes. Insolubilization improved thermal stability of Bam HI and Eco RI. The bound enzyme can be recovered from reaction mixtures and reused several times. Upon storage at 4°C, coupled endonucleases remained stable for several months.

AB - Type II restriction endonucleases Bam HI and Eco RI were covalently coupled to Sepharose. These insolubilized enzymes generated fragment patterns for several viral DNAs identical to those produced by the respective free enzymes. Conditions for optimal activity were similar for both bound and unbound forms of the enzymes. Insolubilization improved thermal stability of Bam HI and Eco RI. The bound enzyme can be recovered from reaction mixtures and reused several times. Upon storage at 4°C, coupled endonucleases remained stable for several months.

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Preparation and properties of insolubilized restriction endonucleases

Abstract

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