PUBLICATION
Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase
- Authors
- Chang, W.N., Tsai, J.N., Chen, B.H., and Fu T.F.
- ID
- ZDB-PUB-051101-6
- Date
- 2006
- Source
- Protein Expression and Purification 46(2): 212-220 (Journal)
- Registered Authors
- Chang, Wen-Ni
- Keywords
- Serine hydroxymethyltransferase, Folate, One-carbon metabolism, B6 enzyme, Zebrafish
- MeSH Terms
-
- Animals
- Cytoplasm/enzymology
- Cytoplasm/genetics
- Escherichia coli/genetics
- Escherichia coli/growth & development
- Gene Expression
- Glycine Hydroxymethyltransferase/biosynthesis*
- Glycine Hydroxymethyltransferase/chemistry
- Glycine Hydroxymethyltransferase/genetics
- Glycine Hydroxymethyltransferase/isolation & purification*
- Humans
- Rabbits
- Recombinant Proteins/biosynthesis*
- Recombinant Proteins/chemistry
- Recombinant Proteins/genetics
- Recombinant Proteins/isolation & purification*
- Sequence Homology, Amino Acid
- Zebrafish/genetics
- Zebrafish Proteins/biosynthesis*
- Zebrafish Proteins/chemistry
- Zebrafish Proteins/genetics
- Zebrafish Proteins/isolation & purification*
- PubMed
- 16242955 Full text @ Protein Expr. Purif.
Citation
Chang, W.N., Tsai, J.N., Chen, B.H., and Fu T.F. (2006) Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase. Protein Expression and Purification. 46(2):212-220.
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 15mg 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping