Abstract
This chapter essentially combines a variety of techniques already described in other chapters to allow a rapid characterization or con firmation of λ expression clones. Antibodies used to screen expres sion libraries (see Chapter 21) may have been raised against affinity purified antigen (see Chapter 29), which can never be guaranteed free of contaminating material, or may even have been produced by an infected host during the course of a parasitic infection. Obviously such sera may recognize more than one antigen, and it is essential for the char acterization of the gene that has been cloned using such sera that the antibodies and gene product generating the positive signal can be identified.
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References
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Ridley, R. G., Patel, H. V., Parfett, C. L. J., Reichhng, S., Olynyk, K. A., and Freeman, K. B. (1986) Immunological detection of cDNA clones encoding the uncoupling protein of brown adipose tissue. Evidence for an antigenic determ inant within the C-terminal eleven amino acids. Bioscience Reports 6, 87–94.
Harlow, E. and Lane, D (1988) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Stahl, H. D., Bianco, A. E, Crewther, P. E., Anders, R. F., Kyne, A. P., Coppel, R L., Mitchell, G. F. Kemp, D. J., and Brown, G. V. (1986) Sorting large num bers of clones expressing Plasmodium falciparum antigens in Escherichia coli by differential antibody screening. Mol. Biol. Med. 3, 351–368.
Hall, R, Hyde, J. E., Goman, M, Simmons, D. L., Hope, I. A., Mackay, M., Scaife, J, Merkli, B., Richie, R., and Stocker, J. (1984) Major surface antigen gene of a human malaria parasite cloned and expressed in bacteria, Nature 311, 379–382.
Ozaki, L. S., Mattei, D., Jendoubi, M, Druilhe, P, Bhsnick, T., Guilotte, M., Puijalon, O., and Pereira da Silva, L. (1986) Plaque antibody selection: rapid immunological analysis of a large number of recombinant phage clones posi tive to sera raised against Plasmodium falciparum antigens. J. Immunol. Methods 89, 213–219.
lams, K. P., Hall, R, Webster, P, and Musoke, A. J. (1990) Identification of λ.gtl 1 clones encoding the major antigenic determinants expressed by Theileria parva sporozoites. Infect. Immun. 58, 1828–1834
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© 1993 Humana Press Inc, Totowa, NJ
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Robert, G., Roger, H. (1993). Antibody Select Procedure for Characterization of Expression Clones. In: Hyde, J.E. (eds) Protocols in Molecular Parasitology. Methods in Molecular Biology™, vol 21. Humana Press. https://doi.org/10.1385/0-89603-239-6:297
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DOI: https://doi.org/10.1385/0-89603-239-6:297
Publisher Name: Humana Press
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