Screening cDNA expression libraries in lambda gt11 with a T cell hybridoma.
A T cell hybridoma specific for a protein of Plasmodium chabaudi chabaudi has been used to test a system for direct T cell screening of a cDNA library of P. chabaudi in the phage lambda gt11. The technique is based upon the rapid separation of the recombinant beta-galactosidase fusion protein from the bacterial mixtures using polystyrene beads coated with anti-beta-galactosidase antibodies. These coated beads are cultured with antigen-presenting cells and the T cell hybridoma. The technique is sufficiently sensitive to pick up the products of one recombinant phage in a pool of 1000-10,000 other phages. Individual plaques or clones of recombinant phage can be selected after testing of sequential dilutions of pools containing positive recombinant phages. This technique will be generally applicable and should be useful for the identification of important T cell peptides in infectious diseases.
Gillard-Blaas
S
S
Max-Planck-Institut für Immunobiologie, Freiburg, F.R.G.
Ayane
M
M
Wirbelauer
C
C
Langhorne
J
J
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D010582:Bacteriophage lambda; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D015723:Gene Library; D005822:Genetic Vectors; D006825:Hybridomas / Q000276:immunology*; D007840:Latex; D051379:Mice; D008815:Mice, Inbred Strains; D008863:Microspheres; D010961:Plasmodium / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D015183:Restriction Mapping; D013601:T-Lymphocytes / Q000276:immunology*
D000953:Antigens, Protozoan; D004274:DNA, Recombinant; D007840:Latex; D011993:Recombinant Fusion Proteins