Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase

Wen Ni Chang; Jen Ning Tsai; Bing Hung Chen; Tzu Fun Fu

doi:10.1007/11816171_28

Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase

Wen Ni Chang, Jen Ning Tsai, Bing Hung Chen, Tzu Fun Fu

Department of Medical Laboratory Science and Biotechnology

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17 Citations (Scopus)

Abstract

A cDNA which encodes for zebrafish serine hydroxymethyltransferase (SHMT) has been cloned into a pET43.1a vector as a NdeI-EcoRI insert and transformed into HMS174(DE3) cells. After induction with isopropyl thiogalactoside, the enzyme was purified with a three-step purification protocol and about 15 mg of pure enzyme was obtained per liter of culture. Spectral and structural characteristics of the recombinant zebrafish SHMT are similar to the rabbit and human cytosolic SHMT. Kinetic constants for the natural substrates l-serine and tetrahydrofolate are also comparable to the values obtained previously for the rabbit and human cytosolic enzyme.

Original language	English
Pages (from-to)	212-220
Number of pages	9
Journal	Protein Expression and Purification
Volume	46
Issue number	2
DOIs	https://doi.org/10.1007/11816171_28
Publication status	Published - 2006 Apr

All Science Journal Classification (ASJC) codes

Biotechnology

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abstract = "A cDNA which encodes for zebrafish serine hydroxymethyltransferase (SHMT) has been cloned into a pET43.1a vector as a NdeI-EcoRI insert and transformed into HMS174(DE3) cells. After induction with isopropyl thiogalactoside, the enzyme was purified with a three-step purification protocol and about 15 mg of pure enzyme was obtained per liter of culture. Spectral and structural characteristics of the recombinant zebrafish SHMT are similar to the rabbit and human cytosolic SHMT. Kinetic constants for the natural substrates l-serine and tetrahydrofolate are also comparable to the values obtained previously for the rabbit and human cytosolic enzyme.",

author = "Chang, {Wen Ni} and Tsai, {Jen Ning} and Chen, {Bing Hung} and Fu, {Tzu Fun}",

note = "Funding Information: This work was supported by Grant NSC 93-2320-B-006-096 from the National Science Council, Taiwan. Our appreciation also goes to Dr. Verne Schirch/Virginia Commonwealth University for his valuable advice and assistance.",

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T1 - Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase

AU - Chang, Wen Ni

AU - Tsai, Jen Ning

AU - Chen, Bing Hung

AU - Fu, Tzu Fun

N1 - Funding Information: This work was supported by Grant NSC 93-2320-B-006-096 from the National Science Council, Taiwan. Our appreciation also goes to Dr. Verne Schirch/Virginia Commonwealth University for his valuable advice and assistance.

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Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase

Abstract

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