Browse > Article

Molecular Cloning and Expression of Rice Lectin in Escherichia coli  

홍성관 (중앙대학교 자연과학대학 화학과)
전상훈 (중앙대학교 자연과학대학 화학과)
김하형 (중앙대학교 약학대학)
공광훈 (중앙대학교 자연과학대학 화학과)
Publication Information
YAKHAK HOEJI / v.46, no.4, 2002 , pp. 270-275 More about this Journal
Abstract
The lectin gene from rice was amplified by the polymerase chain reaction. The amplified DNA was inserted into the expression vector pET26b and expressed it as a fusion protein with polyhistidine sequences in Escherichia coli. The recombinant protein was produced by induction with 0.4 mM isopropyl-$\beta$-D-thiogalactopyranoside at 37$^{\circ}C$ and purified by an immobilized metal affinity chromatography. The recombinant protein was found to have lectin activity by the hemagglutination inhibition assay. The hemagglutination activity of the recombinant protein was optimal at pH 4.0-7.0 and was dependent on $Ca^{2+}$ and Mn$^{2+}$.2+/.
Keywords
Lectin; rice; gene cloning; expression in E. coli;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Rudiger H. : Plant lectins more thanjust tools for glycoscientists; occurrence, structure, and possible functions of plant lectins. Acta. Anat. (Basel), 161, 130 (1998)
2 Gabius, H. J. : Animal lectins. Eur. J. Biochem., 243, 543 (1997)
3 Sharon N., and Lis H. : A century of lectin research (1888-1988) Trends Biochem. Sci., 12, 488 (1987)
4 Miarons, P. B. and Fresno, M. : Lectins from tropical sponges: Purification and characterization of lectins from Genus aplysina. J Biol. Chem., 275, 29283 (2000)
5 Rini J. M. : Lectin structure. Annu. Rev. Biophys. Biomole. Struct., 24, 551 (1995)   DOI   ScienceOn
6 Bradford, M. M. : A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248 (1976)
7 Van Damme, E. J., Leuven, F. V. and Peumans W. J.: Isolation, characterization and molecular cloning of the bark lectins from Maackia amurensis. Glycoconjugate J., 14, 449 (1997)
8 Arango, R., Adar, R., Rozenblatt, S. and Sharon, N. : Expression of Erythrina corallodendron lectin in Escherichia coli. Eur. J. Biochem., 205, 575 (1992)
9 Silva-Lucca, R. A., Tabak, M., Nascimento, O. R., Roque-Barreira, M. C. and Beltramini, L. M. : Structural and thermodynamic studies of $KM^+$, a D-mannose binding lectin from Artocarpus integrifolia seeds. Biophys. Chem., 79, 81 (1999)
10 Van Damme, E. J. M., Peumans, W. J., Barre, A. and Rouge, P.: Plants lectins: A composite of several distinct families of structurally and evolutionary related proteins with diverse biological roles. Crit. Rev. Plant Sci., 17, 575 (1998)
11 Peumans W. J., and Van Damme E. J. : The role of lectins in plant defence. Histochem. J., 27, 253 (1995)
12 Goldstein, I. J., Hughes, R C., Monsigny, M., Osawa, T. and Sharon, N. : What should be called a lectin? Nature, 285, 66 (1980)
13 Weis W. I. : Structural basis of lectin-carbohydrate recognition. Annu. Rev. Biochem., 65, 441 (1996)
14 Laemmli, U. K. : Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680 (1970)
15 Elgavis, S. and Shanan, B. : Lectin-carbohydrate interactions; different folds, common recognition principles. Trends Biochem. Sci., 22, 462 (1997)
16 Adar, R. and Sharon, N. : Mutational studies of the amino acid residues in the combining site of Erythrina corallodendron lectin. Eur. J. Biochem., 239, 668 (1996)
17 Sharon N. : Lectin-carbohydrate complexes of plants and animals: an atomic view. Tends Biochem. Sci., 18, 221 (1993)
18 Lord J. M., Hartley M. R., and Roberts L. M. : Ribosome inactivating proteins of plants. Semin. Cell Biol., 2, 15 (1991)