Abstract
Any biochemical analysis is usually made easier by the availability of large numbers of cells to be analyzed, and one of the reasons for the position held by Trypanusoma brucei as the best characterized parasite is the relative ease with which it can be cultured in the laboratory. The ability to culture cloned populations derived from individual trypanosomes before and after an antigenic switch is vital in investigations into the mechanism of antigenic variation. Genomic DNA prepared from such cloned populations used to analyze variant specific glycoprotein (VSG) genes by Southern blotting led to the discovery of the genomic rearrangements involved in antigenic variation (1–5).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hoeijmakers, J., Frasch, A, Bernards, A, Borst, P., and Cross, G. (1981) Novel expression linker copies of the genes for variant surface antigens in trypanosomes. Nature 284, 78–80.
Pays, E, van Merrvenne, N., Le Ray, D., and Steiner-t, M. (1981) Gene duplication and transposrtron linked to antigenic varration in Trypanosoma brucei Proc. Natl. Acad. Sci. USA 78 2673–2677.
Young, J., Donelson, J, MaJiwa, P, Shapiro, S, and Williams, R (1982) Analysis of genomic rearrangements associated with two variable antigen genes of Trypanosoma brucei Nucl Acids Res. 10, 803–819.
Pays, E. and Steinert, M (1988) Control of antigen gene expression in African trypanosomes Ann. Rev. Genet. 22 107–126.
Borst, P (1986) Discontmuous transcrrption and antigenic variation in trypanosomes Ann Rev. Biochem 55, 701–732
Lanham, S M. (1968) Separation of trypanosomes from the blood of infected rats and mice using anion-exchangers. Nature 218, 1273–1274.
Lanham, S. M. and Godfrey, D. G. (1970) Isolation of salivarian trypanosomes from man and other mammals using DEAE-cellulose. Exp. Parasitol. 28, 521–534.
Cross, G. A. M. (1975) Identificatron, puriftcatron and properties of clone specific glycoprotem antigens constituting the surface coat of Trypanosoma brucei. Parasitology 71, 393–417.
Miller, E. N. and Turner, M. J. (1981) Analysis of antigenic types appearing in first relapse populattons of clones of Trypanosoma brucei Parasitology 82, 63–80.
Rovis, L. and Dube, D (1981) Studies on the btosynthesis of the varrant surface glycoprotein of Trypanosoma brucei: sequence of glycosylation. Mol. Biochem Parasitol. 4, 77–93.
Ferguson, M. J, Duszenko, M, Lamont, G. S, Overath, P, and Cross, G A M. (1986) Biosynthesis of Trypanosoma brucei variant surface glycoproteins J. Biol. Chem. 261, 356–362.
Bangs, J. D., Hereld, D, Krakow, J., Hart, G. W, and Englund, P T (1985) Rapid processing of the carboxyl terminus of a trypanosome variant surface glycoprotein. J. Biol. Chem. 82, 3207–3211.
Ferguson, M. J. and Cross, G. A. M. (1984) Myristylation of the membrane form of a Trypanosoma brucei variant surface glycoprotem. J Biol. Chem. 259, 3011–3015.
Ferguson, M A. J., Haldar, K, and Cross, G. A M. (1985) Trypanosomu brucei variant surface glycoprotein has a sn-1,2-dimyristyl glycerol membrane anchor at its COOH terminus. J Biol Chem. 260, 4963–49
Büllow R and Overath P (1985) Synthesis of a hydrolase for the membrane form variant surface glycoprotetin is repressed during transformation of Trypanosoma brucei FEBS Lett. 187, 105–110
Büllow, R. and Overath, P (1986) Purification and characterrzation of the membrane-form variant surface glycoprotein hydrolase of Trypanosoma brucei J Biol. Chem 261, 11,918–11,923
Hereld, D., Krakow, J L, Bangs, J. D., Hart, G W, and Englund, P. T. (1986) A phosphohpase C from Trypanosoma brucei which selectively cleaves the glycolipid on the variant surface glycoprotein. J Biol. Chem. 261, 13,813–13,819
Fox, J. A., Druszenko, M., Ferguson, M. A. J, Low, M. G., and Cross, G. A. M (1986) Purificatron and characterization of a novel glycan-phosphatidylinositol specific phospholipase C from Trypanosoma brucei. J. Biol. Chem. 261, 15,767–15,771
Brun, R and Schonenberger, M (1979) Cultivation and in vitro cloning of procyclic culture forms of Trypanosoma brucei in a semi-defined medium Acta Trop. 36, 289–292.
Bulow R., Nonnengässer C, and Overath P. (1989) The release of the variable surface glycoprotein on transformation of Trypanosoma brucei from bloodstream to procyclic form. Mol. Biochem. Parantol. 32, 85–92
Harlow, E. and Lane, D (1988) Antibodies, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Doering, T., Masterson, W, Englund, P., and Hart, G. (1989) Biosynthesis of the glycosyl phosphatidylinositol of the trypanosome variant surface glycoprotein J. Biol. Chem. 264, 11,168–11,173.
Krakow, J L., Hereld, D, Bangs, J D., Hart, G. W, and Englund, P T (1986) Identification of a glycolipid precursor of the Trypanosoma brucei variant surface glycoprotein. J. Biol. Chem. 261, 12,147–12,153.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1993 Humana Press Inc, Totowa, NJ
About this protocol
Cite this protocol
Carrington, M. (1993). Culturing and Biological Cloning of Trypanosoma brucei. In: Hyde, J.E. (eds) Protocols in Molecular Parasitology. Methods in Molecular Biology™, vol 21. Humana Press. https://doi.org/10.1385/0-89603-239-6:1
Download citation
DOI: https://doi.org/10.1385/0-89603-239-6:1
Publisher Name: Humana Press
Print ISBN: 978-0-89603-239-2
Online ISBN: 978-1-59259-508-2
eBook Packages: Springer Protocols