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Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase.
Protein Expr Purif. 2006 Apr; 46(2):212-20.PE

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.

Authors+Show Affiliations

Department of Medical Laboratory Science and Biotechnology, National Cheng Kung University, School of Medicine, Tainan 701, Taiwan.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16242955

Citation

Chang, Wen-Ni, et al. "Cloning, Expression, Purification, and Characterization of Zebrafish Cytosolic Serine Hydroxymethyltransferase." Protein Expression and Purification, vol. 46, no. 2, 2006, pp. 212-20.
Chang WN, Tsai JN, Chen BH, et al. Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase. Protein Expr Purif. 2006;46(2):212-20.
Chang, W. N., Tsai, J. N., Chen, B. H., & Fu, T. F. (2006). Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase. Protein Expression and Purification, 46(2), 212-20.
Chang WN, et al. Cloning, Expression, Purification, and Characterization of Zebrafish Cytosolic Serine Hydroxymethyltransferase. Protein Expr Purif. 2006;46(2):212-20. PubMed PMID: 16242955.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR 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, Y1 - 2005/09/28/ PY - 2005/06/21/received PY - 2005/08/10/revised PY - 2005/08/16/accepted PY - 2005/10/26/pubmed PY - 2006/5/31/medline PY - 2005/10/26/entrez SP - 212 EP - 20 JF - Protein expression and purification JO - Protein Expr. Purif. VL - 46 IS - 2 N2 - 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. SN - 1046-5928 UR - https://www.unboundmedicine.com/medline/citation/16242955/Cloning_expression_purification_and_characterization_of_zebrafish_cytosolic_serine_hydroxymethyltransferase_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1046-5928(05)00285-8 DB - PRIME DP - Unbound Medicine ER -