Cloning, expression, and purification of recombinant Neisseria gonorrhoeae proteins
Cloning, expression, and purification of recombinant Neisseria gonorrhoeae proteins
Modern DNA recombinant techniques and major advances in genetic engineering have resulted in the development of bacterial expression systems that guarantee an unlimited supply of valuable proteins that have potential clinical or industrial use, but which are often limited by their low natural availability. This chapter provides the reader with a general scheme to clone, express, and purify native histidine (His)-tagged proteins in the desired quantity and quality required for its intended use, and reviews the most important factors affecting the production of recombinant proteins in a soluble form. Alternative methods for purification of insoluble recombinant proteins under denaturing conditions are also discussed. An optimized protocol to successfully purify native Neisseria gonorrhoeae Adhesin Complex Protein (Ng-ACP; NGO1981) is used as a technical example for the processes, which could potentially be applied to any gonococcal recombinant protein of interest.
Affinity chromatography, Cloning, His-tag, Neisseria gonorrhoeae, Ng-ACP (NGO1981), Recombinant protein purification
233-266
Humbert, María Victoria
82134d25-24b8-4fdd-bd1c-461683b5322e
2019
Humbert, María Victoria
82134d25-24b8-4fdd-bd1c-461683b5322e
Humbert, María Victoria
(2019)
Cloning, expression, and purification of recombinant Neisseria gonorrhoeae proteins.
In,
Christodoulides, M.
(ed.)
Methods in Molecular Biology.
(Methods in Molecular Biology, 1997)
New York.
Humana, .
(doi:10.1007/978-1-4939-9496-0_15).
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Book Section
Abstract
Modern DNA recombinant techniques and major advances in genetic engineering have resulted in the development of bacterial expression systems that guarantee an unlimited supply of valuable proteins that have potential clinical or industrial use, but which are often limited by their low natural availability. This chapter provides the reader with a general scheme to clone, express, and purify native histidine (His)-tagged proteins in the desired quantity and quality required for its intended use, and reviews the most important factors affecting the production of recombinant proteins in a soluble form. Alternative methods for purification of insoluble recombinant proteins under denaturing conditions are also discussed. An optimized protocol to successfully purify native Neisseria gonorrhoeae Adhesin Complex Protein (Ng-ACP; NGO1981) is used as a technical example for the processes, which could potentially be applied to any gonococcal recombinant protein of interest.
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e-pub ahead of print date: 23 May 2019
Published date: 2019
Keywords:
Affinity chromatography, Cloning, His-tag, Neisseria gonorrhoeae, Ng-ACP (NGO1981), Recombinant protein purification
Identifiers
Local EPrints ID: 433394
URI: http://eprints.soton.ac.uk/id/eprint/433394
ISSN: 1064-3745
PURE UUID: 86910867-afd5-4675-9b53-98e68850abfa
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Date deposited: 19 Aug 2019 16:30
Last modified: 16 Mar 2024 04:19
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Author:
María Victoria Humbert
Editor:
M. Christodoulides
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