Full Text:   <2856>

CLC number: Q555+.4

On-line Access: 2024-08-27

Received: 2023-10-17

Revision Accepted: 2024-05-08

Crosschecked: 0000-00-00

Cited: 0

Clicked: 5291

Citations:  Bibtex RefMan EndNote GB/T7714

-   Go to

Article info.
1. Reference List
Open peer comments

Journal of Zhejiang University SCIENCE A 2004 Vol.5 No.2 P.164-172

http://doi.org/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


Share this article to: More

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 A, 2004, 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 A",
volume="5",
number="2",
pages="164-172",
year="2004",
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 A
%V 5
%N 2
%P 164-172
%@ 1869-1951
%D 2004
%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 A
VL - 5
IS - 2
SP - 164
EP - 172
%@ 1869-1951
Y1 - 2004
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.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1] Akimoto, Y., Obinata, A., Endo, H., Furukawa, K., Aoki, D., Nozawa, S., Hirano, H., 1995. Immunocytochemical localization of the protein reactive to human beta-1,4-galactosyltransferase antibodies during chick embryonic skin differentiation.Anat. Rec.,243(1):109-119.

[2] Almeida, R., Amado, M., David, D., Levery, S.B., Holmes, E.H., Merkx, G., van Kessel, A.G., Rygaard, E., Hassan, H., Bennett, E., Clausen, H., 1997. A Family of humanβ4-galactosyltransferases: cloning and expression of two novel UDP-galactoseβ-N-acetyl-glucosamineβ-1,4-gal-actosyltransferases:β4-Gal-T2 andβ4Gal-T3.J. Biol. Chem.,272(51):31979-31991.

[3] Appeddu, P.A., Shur, B.D., 1994. Molecular analysis of cell surface beta-1,4-galactosyltransferase function during cell migration.Proc. Natl. Acad. Sci. U.S.A,91(6):2095-2099.

[4] Asano, M., Furukawa, K., Kido, M., Matsumoto, S., Umesaki, Y., Kochibe, N., Iwakura, Y., 1997. Growth retardation and early death ofβ-1,4-galactosyltransferase knockout mice with augmented proliferation and abnormal differentiation of epithelial cells.EMBO J.,16(8):1850-1857.

[5] Axford, J.S., 1999. Glycosylation and rheumatic disease.Biochim. Biophys. Acta.,1455(2-3):219-229.

[6] Bayna, E.M., Shaper, J.H., Shur, B.D., 1988. Temprally specific involvement of cell surfaceβ-1,4-galactosyl-transferase during mouse embryo morula compaction.Cell,53:145-157.

[7] Begovac, P.C., Shur, B.D., 1990. Cell surface galactosyltransferase mediates the initiation of neurite out-growth from PC 12 cells on laminin.J. Cell Biol.,110(2):461-470.

[8] Eckstein, D.J., Shur, B.D., 1992. Cell surface beta-1,4-galactosyltransferase is associated with the detergent-insoluble cytoskeleton on migrating mesenchymal cells.Exp. Cell Res.,201(1):83-90.

[9] Evans, S.C., Youakim, A., Shur, B.D., 1995. Biological consequences of targetingβ-1,4-galactosyltransferase to two different subcellular compartments.Bioessays,17(3):261-268.

[10] Fan, Y., Yu, L., Tu, Q., Gong, R., Jiang, Y., Zhang, Q., Dai, F., Chen, C., Zhao, S., 2002. Molecular cloning, genomic organization, and mapping ofβ4GalT-VIb, a brain abundant member ofβ4-galactosyltransferase gene family, to human chromosome 18q12.1.DNA Seq.,13(1):1-8.

[11] Gillespie, P.G., Hudspeth, A.J., 1991. High-purity isolation of bullfrog hair bundles and subcellular and topological localization of constituent proteins.J. Cell Biol.,112(4):625-640.

[12] Hathaway, H.J., Shur, B.D., 1992. Cell surface beta-1,4-galactosyltransferase functions during neural crest cell migration and neurulationin vivo.J. Cell Biol.,117(2):369-382.

[13] Hathaway, H.J., Shur, B.D., 1996. Mammary gland morphogenesis is inhibited in transgenic mice that overexpress cell surface beta-1,4-galactosyltranferase.Development,122(9):2859-2872.

[14] Holm, L., Sander, C., 1983. Protein structure comparison by alignment of distance matrices.J. Mol. Biol.,233:123-138.

[15] Huang, Q., Shur, B.D., Begovac, P.C., 1995. Overexpressing cell surface beta-1,4-galactosyltransferase in PC12 cells increases neurite outgrowth on laminin.J. Cell Sci.,108(Pt 2):839-847.

[16] Lopez, L.C., Youakim, A., Evans, S.C., Shur, B.D., 1991. Evidence for a molecular distinction between Golgi and cell surface forms ofβ-1,4-galactosytransferase.J. Bio. Chem.,266(24):15984-15991.

[17] Macek, M.B., Lopez, L.C., Shur, B.D., 1991. Aggregation of beta-1,4-galactosyltransferase on mouse sperm induces the acrosome reaction.Dev. Biol.,147(2):440-444.

[18] Maillet, C.M., Shur, B.D., 1993. Uvomorulin, LAMP-1, and laminin are substrates for cell surface beta-1,4-galactosyltransferase on F9 embryonal carcinoma cells: comparisons between wild-type and mutant 5.51 att-cells.Exp. Cell Res.,208(1):282-295.

[19] Maillet, C.M., Shur, B.D., 1994. Perturbing cell surface beta-(1,4)-galactosyltransferase on F9 embryonal carcinoma cells arrests cell growth and induces laminin synthesis.Cell Sci.,107(Pt 6):1713-1724.

[20] Malissard, M., Zeng, S., Berger, E.G., 2000. Expression of functional soluble forms of humanβ-1,4-galactosyltransferase I, α-2,6-sialyltransferase, and α-1,3-fucosyltransferase VI in the methylotrophic yeast Pichia pastoris.Biochem. Biophys. Res. Commun.,267(1):169-173.

[21] Malissard, M., Berger, E.G., 2001. Improving solubility of catalytic domain of human beta-1,4-galactosyltransferase 1 through rationally designed amino acid replacements.Eur. J. Biochem.,268(15):4352-4358.

[22] Masibay, A.S., Balaji, P.V., Boeggeman, E.E., Qasba, P.K., 1993. Mutational analysis of the golgi retention signal of bovineβ-1,4-galactosyltransferase.J. Bio. Chem.,268(13):9908-9916.

[23] Masri, K.A., Appert, H.E., Fukuda, M.N., 1988. Identification of the full-length coding sequence of human galactosylstransferase(β-N-acetylglucosminide:β-1,4-galactosyltransferase).Biochem. Biophys. Res. Commun.,157(2):657-663.

[24] McDonald, O.B., Chen, W.J., Ellis, B., Hoffman, C., Overton, L., Rink, M., Smith, A., Marshall, C.J., Wood, E.R., 1999. A scintillation proximity assay for the Raf/MEK/ERK kinase cascade: high-throughput scrceening and identification of selective enzyme inhibitors.Anal. Biochem.,268(2):318-329.

[25] Mengle, G.L., Mccoy, H.M.F., Tiemeier, D.C., 1991. Genomic structure and expression of human beta-1,4-galactosyltransferase.Biochem. Biophy. Res. Commun.,176:1269-1276.

[26] Nomura, T., Takizawa, M., Aoki, J., Arai, H., Inoue, K., Wakisaka, E., Yoshizuka, N., Imokawa, G., Dohmae, N., Takio, K., Hattori, M., Matsuo, N., 1988. Purification, cDNA cloning, and expression of the UDP-Gal: glucosylceramideβ-1,4-galactosyltransferase from rat brain.J. Biol. Chem.,273(22):13570-13577.

[27] Park, J.E., Lee, K.Y., Do, S.I., Lee, S.S., 2002. Expression and characterization ofβ-1,4-galactosyltransferase fromNeisseria meningitisdisandNeisseria gonorrhoeae.J. Biochem. Mol. Biol.,35(3):330-336.

[28] Posern, G., Zheng, J., Knudsen, B.S., Kardinal, C., Muller, K.B., Voss, J., Shishido, T., Cowburn, D., Cheng, G., Wang, B., Kruh, G.D., Burrell, S.K., Jacobson, C.A., Lenz. D.M., Zamborelli, T.J., Adermann, K., Hanafusa, H., Feller, S.M., 1998. Development of highly selective SH3 binding peptides for Crk and CRKL which disrupt Crk-complexes with DOCK180, SoS and C3G.Oncogene,16(15):1903-1912.

[29] Sambrook, J., Fritsh, E.F., Maniatis, T., 1989. Molecular Cloning: A Laboratory Manual, 2nd Edition. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

[30] Shi, X., Amindari, S., Paruchuru, K., Skalla, D., Burkin, H., Shur, B.D., Miller, D.J., 2001. Cell surface beta-1,4-galactosyltransferase-I activates G protein-dependent exocytotic signaling.Development,128(5):645-654.

[31] Smith, D.B., Johnson, K.S., 1988. Single-step purifycation of polypeptides expressed inEscherichia colias fusions with glutathione S-transferase.Gene,67(1):31-40.

[32] Tomiya, N., Howe, D., Aumiller, J.J., Pathak, M., Park, J., Palter, K.B., Jarvis, D.L., Betenbaugh, M.J., Lee, Y.C., 2003. Complex-type biantennary N-glycans of recombinant human transferrin fromTrichoplusia niinsect cells expressing mammalian {beta}-1,4-galactosyltrans-ferase and {beta}-1,2-N-acetylgluco-saminyltransferase II.Glycobiology,13(1):23-34.

[33] Uehara, K., 1998. Structural analysis of milk and testisβ-1,4-galactosyltransferase gene products.Cell structure and function,23:43-48.

[34] Vadaie, N., Hulinsky, R.S., Jarvis, D.L., 2002. Identification and characterization of aDrosophila melanogasterortholog of humanβ-1,4-galactosyltransferase VII.Glycobiology,12(10):589-597.

[35] Vrielink, A., Rüger, W., Driessen, H.P., Freemont, P.S., 1994. Crystal structure of the DNA modifying enzymeβ-glucosyltransferase in the presence or in the absence of the substrate uridine diphosphoglucose.EMBO J.,13(15):3413-3422.

[36] Wassler, M.J., Shur, B.D., 2000. Clustering of cell surface beta-1,4-galactosyltransferase I induces transient tyrosine phosphorylation of focal adhesion kinase and loss of stress fibers.J. Cell Sci.,113(Pt 2):237-245.

[37] Wassler, M.J., Foote, C.J., Gelman, I.H., Shur, B.D., 2001. Functional interaction between the SSeCKS scaffolding protein and the cytoplasmic domain of beta-1,4-galactosyltransferase.J. Cell Sci.,114:2291-2300.

[38] Yang, R.Y., Hsu, D.K., Liu, F.T., 1996. Expression of galectin-3 modulates T-cell growth and apoptosis.Proc. Natl. Acad. Sci. U.S.A,93(13):6737-6742.

[39] Youakim A., Hathaway H.J., Miller, D.J., Gong, X., Shur, B.D., 1994. Overexpressing sperm surfaceβ1,4-galactosyltransferase in transgenic mice affects multiple aspects of sperm-egg interactions.J. Cell Biol.,126(6):1573-1583.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE