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Literature DB >> 2583511

Purification and characterization of the FokI restriction endonuclease.

T Kaczorowski¹, P Skowron, A J Podhajska.

Abstract

The restriction endonuclease FokI from Flavobacterium okeanokoites was purified to homogeneity. Based on gel filtration, sedimentation and sodium dodecyl sulfate-polyacrylamide-gel electrophoresis, the following properties of the enzyme were determined: FokI exists in one active monomeric form, and has an Mr of 64-65.4 x 10(3).FokI is a strongly basic protein with an isoelectric point of 9.4. The enzyme exhibits restriction activity in the pH range 5.0 to 10.5 (maximum level at pH 7.0-8.5) and its divalent cation requirement is satisfied not only by Mg2+, but also by Co2+, Mn2+, Ni2+, Cd2+, Zn2+ and Fe2+.

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Year: 1989 PMID： 2583511 DOI： 10.1016/0378-1119(89)90285-0

Source DB: PubMed Journal: Gene ISSN： 0378-1119 Impact factor: 3.688

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Cited

17 in total

1. Engineering a nicking endonuclease N.AlwI by domain swapping.

Authors: Y Xu; K D Lunnen; H Kong
Journal: Proc Natl Acad Sci U S A Date: 2001-10-30 Impact factor: 11.205

2. The nicking endonuclease N.BstNBI is closely related to type IIs restriction endonucleases MlyI and PleI.

Authors: L S Higgins; C Besnier; H Kong
Journal: Nucleic Acids Res Date: 2001-06-15 Impact factor: 16.971

3. The FokI methyltransferase from Flavobacterium okeanokoites. Purification and characterization of the enzyme and its truncated derivatives.

Authors: T Kaczorowski; M Sektas; P Skowron; A J Podhajska
Journal: Mol Biotechnol Date: 1999-11 Impact factor: 2.695

4. A new Thermus sp. class-IIS enzyme sub-family: isolation of a 'twin' endonuclease TspDTI with a novel specificity 5'-ATGAA(N(11/9))-3', related to TspGWI, TaqII and Tth111II.

Authors: Piotr M Skowron; Jarosław Majewski; Agnieszka Zylicz-Stachula; Sylwia M Rutkowska; Izabela Jaworowska; Renata I Harasimowicz-Słowińska
Journal: Nucleic Acids Res Date: 2003-07-15 Impact factor: 16.971

10. Genetic organization and molecular analysis of the EcoVIII restriction-modification system of Escherichia coli E1585-68 and its comparison with isospecific homologs.

Authors: Iwona Mruk; Tadeusz Kaczorowski
Journal: Appl Environ Microbiol Date: 2003-05 Impact factor: 4.792

Purification and characterization of the FokI restriction endonuclease.

1. Engineering a nicking endonuclease N.AlwI by domain swapping.

2. The nicking endonuclease N.BstNBI is closely related to type IIs restriction endonucleases MlyI and PleI.

3. The FokI methyltransferase from Flavobacterium okeanokoites. Purification and characterization of the enzyme and its truncated derivatives.

4. A new Thermus sp. class-IIS enzyme sub-family: isolation of a 'twin' endonuclease TspDTI with a novel specificity 5'-ATGAA(N(11/9))-3', related to TspGWI, TaqII and Tth111II.

5. Inhibition of DNA replication of human papillomavirus by using zinc finger-single-chain FokI dimer hybrid.

6. FokI dimerization is required for DNA cleavage.

7. Functional domains in Fok I restriction endonuclease.

8. Single amino acid substitutions uncouple the DNA binding and strand scission activities of Fok I endonuclease.

9. Alteration of the cleavage distance of Fok I restriction endonuclease by insertion mutagenesis.

10. Genetic organization and molecular analysis of the EcoVIII restriction-modification system of Escherichia coli E1585-68 and its comparison with isospecific homologs.