Please use this identifier to cite or link to this item: http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/7773
Title: Heterologous Expression and High Degree Purification of the Restriction Endonuclease SauUSI
Authors: TUMULURI, VINAYAK SADASIVAM
SAIKRISHNAN, KAYARAT
Dept. of Biology
Keywords: Restriction endonuclease
Heterologous expression
Affinity chromatography
Anion exchange chromatography
Size exclusion chromatography
2022
Issue Date: Jan-2022
Publisher: Bio-protocol, LLC
Citation: Bio-protocol, 12(1).
Abstract: Mechanisms that target and destroy foreign nucleic acids are major barriers to horizontal gene transfer (HGT) in prokaryotes. Amongst them, restriction-modification (R-M) systems are found in ≥75% of the sequenced genomes in Bacteria and Archaea. Due to their high target sequence specificity and potent nucleolytic activity, R-M systems are used as a paradigm to elucidate the mechanisms of DNA binding and cleavage. Since these enzymes modulate HGT, they are one of the machineries implicated in the ability of a bacterium to gain antibiotic resistance. This protocol provides a detailed purification strategy for the Type IV restriction endonuclease SauUSI from Staphylococcus aureus. This protocol eventually leads to ≥95% purity of protein which can then be used for crystallographic and biochemical purposes.
URI: https://doi.org/10.21769/BioProtoc.4275
http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/7773
ISSN: 2331-8325
Appears in Collections:JOURNAL ARTICLES

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.