| Bioscience & Biotechnology |
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| Cloning and GST-fused expression in E. coli of mouse β-1,4-galactosyltransferase |
| GONG Xing-guo, ZHONG Wen-tao, WU Wen-ying |
| Institute of Biomacromolecule & Enzyme Engineering, College of Life Sciences, Zhejiang University, Hangzhou 310027, China |
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Abstract 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 isopropyl-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.
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Received: 16 January 2003
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