Production of a functional dihydrofolate synthase with a cleavable poly-His tag in Saccharomyces cerevisiae

Onisha Patel; Laura A Castelli; Ross Fernley; Peter Coloe; Ian Macreadie

doi:10.1023/A:1015051526401

Production of a functional dihydrofolate synthase with a cleavable poly-His tag in Saccharomyces cerevisiae

Onisha Patel, Laura A Castelli, Ross Fernley, Peter Coloe, Ian Macreadie

Biochemistry & Molecular Biology

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

4 Citations (Scopus)

Abstract

Polyhistidine-tagged dihydrofolate synthase (DHFS) has been produced in the yeast, Saccharomyces cerevisiae, using a Cu2+-inducible expression system. The tagged DHFS is functional in vivo and was purified using immobilised metal affinity chromatography. A linker of a minimal size allows efficient cleavage of the poly-His tag using thrombin. At least 10 mg of pure DHFS can be recovered per litre of culture.

Original language	English
Pages (from-to)	657 - 662
Number of pages	6
Journal	Biotechnology Letters
Volume	24
Issue number	8
DOIs	https://doi.org/10.1023/A:1015051526401
Publication status	Published - 2002

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10.1023/A:1015051526401

Cite this

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title = "Production of a functional dihydrofolate synthase with a cleavable poly-His tag in Saccharomyces cerevisiae",

abstract = "Polyhistidine-tagged dihydrofolate synthase (DHFS) has been produced in the yeast, Saccharomyces cerevisiae, using a Cu2+-inducible expression system. The tagged DHFS is functional in vivo and was purified using immobilised metal affinity chromatography. A linker of a minimal size allows efficient cleavage of the poly-His tag using thrombin. At least 10 mg of pure DHFS can be recovered per litre of culture.",

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AU - Patel, Onisha

AU - Castelli, Laura A

AU - Fernley, Ross

AU - Coloe, Peter

AU - Macreadie, Ian

PY - 2002

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AB - Polyhistidine-tagged dihydrofolate synthase (DHFS) has been produced in the yeast, Saccharomyces cerevisiae, using a Cu2+-inducible expression system. The tagged DHFS is functional in vivo and was purified using immobilised metal affinity chromatography. A linker of a minimal size allows efficient cleavage of the poly-His tag using thrombin. At least 10 mg of pure DHFS can be recovered per litre of culture.

U2 - 10.1023/A:1015051526401

DO - 10.1023/A:1015051526401

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VL - 24

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JO - Biotechnology Letters

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