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Bio-Design and Manufacturing  2018 Vol.5 No.2 P.164~172

10.1631/jzus.2004.0164


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


Author(s):  GONG Xing-guo, ZHONG Wen-tao, WU Wen-ying

Affiliation(s):  Institute of Biomacromolecule & Enzyme Engineering, College of Life Sciences, Zhejiang University, Hangzhou 310027, China

Corresponding email(s):   Gongxg@cls.zju.edu.cn

Key Words:  &beta, -1, 4-galactosyltransferase (&beta, 4Gal-T), Cloning, GST-fusion


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GONG Xing-guo, ZHONG Wen-tao, WU Wen-ying. Cloning and GST-fused expression in E. coli of mouse β-1,4-galactosyltransferase[J]. Journal of Zhejiang University Science D, 2018, 5(2): 164~172.

@article{title="Cloning and GST-fused expression in E. coli of mouse β-1,4-galactosyltransferase",
author="GONG Xing-guo, ZHONG Wen-tao, WU Wen-ying",
journal="Journal of Zhejiang University Science D",
volume="5",
number="2",
pages="164~172",
year="2018",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2004.0164"
}

%0 Journal Article
%T Cloning and GST-fused expression in E. coli of mouse β-1,4-galactosyltransferase
%A GONG Xing-guo
%A ZHONG Wen-tao
%A WU Wen-ying
%J Journal of Zhejiang University SCIENCE D
%V 5
%N 2
%P 164~172
%@ 1869-1951
%D 2018
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2004.0164

TY - JOUR
T1 - Cloning and GST-fused expression in E. coli of mouse β-1,4-galactosyltransferase
A1 - GONG Xing-guo
A1 - ZHONG Wen-tao
A1 - WU Wen-ying
J0 - Journal of Zhejiang University Science D
VL - 5
IS - 2
SP - 164
EP - 172
%@ 1869-1951
Y1 - 2018
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2004.0164


Abstract: 
&beta;-1,4-galactosyltransferase (&beta;4Gal-T) (EC 2.4.1.38) plays a multifunctional role in many aspects of normal cell physiology. By now, several dozens of &beta;4Gal-T genes have been cloned, separated from mouse, chick, bovine, human, etc. This paper presents the cloning and GST-fused expression of mouse &beta;4Gal-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-&beta;-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 &beta;4Gal-T. The transcriptional product of &beta;4Gal-T gene was proved by Western hybridization analysis to be due to GST-fusion.

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