Cloning and GST-fused expression in E. coli of mouse beta-1,4-galactosyltransferase.
Beta-1,4-galactosyltransferase (beta4Gal-T) (EC 2.4.1.38) plays a multifunctional role in many aspects of normal cell physiology. By now, several dozens of beta4Gal-T genes have been cloned, separated from mouse, chick, bovine, human, etc. This paper presents the cloning and GST-fused expression of mouse beta4Gal-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 beta4Gal-T. The transcriptional product of beta4Gal-T gene was proved by Western hybridization analysis to be due to GST-fusion.
Gong
Xing-guo
XG
Institute of Biomacromolecule & Enzyme Engineering, College of Life Sciences, Zhejiang University, Hangzhou 310027, China. Gongxg@cls.zju.edu.cn
Zhong
Wen-tao
WT
Wu
Wen-ying
WY
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009237:N-Acetyllactosamine Synthase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D010802:Phylogeny; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D014162:Transfection / Q000379:methods
D011993:Recombinant Fusion Proteins; D009237:N-Acetyllactosamine Synthase; D005982:Glutathione Transferase