Cloning and GST－fused expression in E. coli of mouse β－1,4－galactosyltransferase

• Published in: Journal of Zhejiang University. Science 2004-02, Vol.5 (2), p.164-172
• Main Author: 龚兴国 钟文涛 吴文英
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Cloning, Molecular - methods; Escherichia coli - enzymology; Escherichia coli - genetics; Gene Expression Regulation, Bacterial - physiology; Gene Expression Regulation, Enzymologic - physiology; Glutathione Transferase - biosynthesis; Glutathione Transferase - genetics; Mice; Molecular Sequence Data; Molecular Weight; N-Acetyllactosamine Synthase - biosynthesis; N-Acetyllactosamine Synthase - chemistry; N-Acetyllactosamine Synthase - genetics; Phylogeny; Recombinant Fusion Proteins - biosynthesis; Sequence Homology, Amino Acid; Transfection - methods; β-1,4-半乳糖转移酶; 克隆技术; 基因表达; 膜蛋白质
• ISSN: 1009-3095
• DOI: 10.1631/jzus.2004.0164

β-1,4-galactosyltransferase (β4Gal-T) (EC 2.4.1.38) plays a multifunctional role in many aspects of normal cell physiology. By now, several dozens of β4Gal-T genes have been cloned, separated from mouse, chick, bovine, human, etc. This paper presents the cloning and GST-fused expression of mouse β4Gal-T gene in Escherichia coli (E. coli). The target gene was cloned by PCR, followed by identification by DNA sequencing and expression in E.coli with isopro-pyl-β-D-thiogalactoside (IPTG) gradient concentrations, products of which were separated on SDS-PAGE showing that the target protein had the same molecular weight as that of mouse β4Gal-T. The transcriptional product of β4Gal-T gene was proved by Western hybridization analysis to be due to GST-fusion.