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

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

Research output: Contribution to journalArticlepeer-review

16 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 languageEnglish
Pages (from-to)212-220
Number of pages9
JournalProtein Expression and Purification
Volume46
Issue number2
DOIs
Publication statusPublished - 2006 Apr

All Science Journal Classification (ASJC) codes

  • Biotechnology

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