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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 |
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