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

  • Bioscience & Biotechnology
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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 inEscherichia coli (E. coli). The target gene was cloned by PCR, followed by identification by DNA sequencing and expression inE.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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Authors and Affiliations

  1. Institute of Biomacromolecule & Enzyme Engineering, College of Life Sciences, Zhejiang University, 310027, Hangzhou, China

    Gong Xing-guo, Zhong Wen-tao & Wu Wen-ying

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  1. Gong Xing-guo
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  2. Zhong Wen-tao
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  3. Wu Wen-ying
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Correspondence to Gong Xing-guo.

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Project (No. 171026) supported by Cao Guangbiao High-Tech Development Funds of Zhejiang University, China

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Xing-guo, G., Wen-tao, Z. & Wen-ying, W. Cloning and GST-fused expression inE. coli of mouse β-1,4-galactosyltransferase. J. Zheijang Univ.-Sci. 5, 164–172 (2004). https://doi.org/10.1631/BF02840918

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  • DOI: https://doi.org/10.1631/BF02840918

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