Abstract
Over the last decade major advances in cloning technology have markedly improved the ability to construct clone libraries expressing foreign proteins and to screen them to identify clones encoding any desired gene product. A number of types of libraries, expression vec- tors, and screening methods have been described but workers in the field of parasitology have generally used antibody screening of cDNA libraries constructed in the bacteriophage Xgtll or its variants. This chapter will largely detail screening methods developed for this sys- tem, an approach that has been successful in identifying clones expres- sing proteins derived from all classes of parasites including protozoa (Plasmodium, Leishmania, Toxoplasma, and Trypanosoma), helminths (Ascaris, Taenia, and Schistosoma), and ectoparasites (ticks and mites).
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References
Huynh, T. V., Young, R. A., and Davis, R. W. (1985) Constructing and screen-ing cDNA libraries in λA,gt10 and λA,gtl 1, in DNA Cloning: A Practical Approach (Glover, D. M., ed.), IRL Press, Oxford, pp. 49–78.
Sambrook, J., Fritsch, E F., and Maniatis, T. (1989) Molecular Cloning-A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 12.4–12.29.
Dame, J B., Williams, J. L., McCutchan, T. F., Weber, J. L., Wirtz, R. A., Hockmeyer, W. T., Maloy, W L., Schneider, I, Roberts, D., Sanders, G. S., Reddy, E. P, Diggs, C L., and Miller, L. H. (1984) Structure of the gene encoding the immunodominant surface antigen on the sporozoite of the human malaria parasite Plasmodium falciparum. Science 225, 593–599.
Young, R. A. and Davis, R W. (1983) Efficient isolation of genes by using antibody probes. Proc. Natl Acad Set. USA 80, 1194–1198.
Kemp, D.J., Coppel, R L., Cowman, A. F., Saint, R. B., Brown, G. V., and Anders, R F.(1983) Expression of Plasmodium falciparum blood-stage anti gens in Escherichia coli. detection with antibodies from immune humans Proc Natl Acad. Sci. USA 80, 3787–3791.
Smith, D.B. and Johnson, K. S.(1988) Single-step purification of polypep tides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67, 31–40
Elliott, J. F., Albrecht, G. R., Gilladoga, A., Handunnetti, S. M., Neequaye, J, Lallinger, G., Minjas, J. N, and Howard, R J. (1990) Genes for Plasmodium falciparum surface antigens cloned by expression in COS cells. Proc. Natl Acad. Sci. USA 87, 6363–6367.
Stahl, H D., Coppel, R. L., Brown, G. V., Saint, R., Lingelbach, K., Cowman, A. F., Anders, R F., and Kemp, D. J.(1984) Differential antibody screening of cloned Plasmodium falciparum sequences expressed inEscherichia coli: pro cedure for isolation of defined antigens and analysis of human antiseraProc. Natl. Acad. Sci USA 81, 2456–2460.
Blake, M. S., Johnston, K.H., Russel-Jones, G. J., and Gotschilch. E C(1984) A rapid, sensitive method for detection of alkaline phosphatase-conjugated antibody on western blots. Anal. Biochem. 136, 175–179.
Mierendorf, R. C, Percy, C, and Young, R. A(1987) Gene isolation by screen ing λX,gtl 1 libraries with antibodies. Meth Enzymol 152, 458–469.
Ausubel, F. M., Brent, R., Kingston, R E., Moore, D. D., Seidman, J. G, Smith, J. A., and Struhl, K(1990) Current Protocols in Molecular Biology.John Wiley and Sons, New York, Section 6.7
Titus, D. E(1991) Promega protocols and applications guide Promega Cor poration, Madison, WI, p 422
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, Druihle, P., Blisnick, T., Guillotte, M, Puijalon, 0., and Pereirada 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.
Coppel, R. L., Cowman, A F., Lingelbach, K. R., Brown, G. V., Saint, R. B, Kemp, D. J., and Anders, R F.(1983) Isolate-specific S-antigen of Plasmodium falciparum contains a repeated sequence of eleven amino acids Nature 306, 751–756
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© 1993 Humana Press Inc, Totowa, NJ
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Coppel, R.L., Smith, F.M., Petersen, C. (1993). Antibody Screening of Expression Libraries. In: Hyde, J.E. (eds) Protocols in Molecular Parasitology. Methods in Molecular Biology™, vol 21. Humana Press. https://doi.org/10.1385/0-89603-239-6:277
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DOI: https://doi.org/10.1385/0-89603-239-6:277
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