Surface display of a parasite antigen in the ciliate Tetrahymena thermophila.
The ciliated protozoan, Tetrahymena thermophila, offers an attractive medium for the expression of heterologous proteins and could prove particularly useful for the display of foreign proteins on the cell surface. Although progress has been made in transformation of Tetrahymena with heterologous DNA, methods that permit reliable expression of foreign genes have been lacking. Using a mutant strain of T. thermophila carrying a negatively selectable allele of a beta-tubulin gene, we have been able to direct foreign genes to this locus by homologous recombination. Transformed cell lines producing foreign proteins were readily identified and, in at least one case, targeting of proteins to the plasma membrane was accomplished.
Gaertig
J
J
Department of Cellular Biology, University of Georgia, Athens 30602, USA. jgaertig@cb.uga.edu
Gao
Y
Y
Tishgarten
T
T
Clark
T G
TG
Dickerson
H W
HW
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000378:metabolism; D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism; D016798:Ciliophora / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D004351:Drug Resistance / Q000235:genetics; D005393:Fish Diseases / Q000469:parasitology; D015870:Gene Expression; D015151:Immunoblotting; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D017239:Paclitaxel / Q000494:pharmacology; D015800:Protozoan Proteins; D011995:Recombination, Genetic; D016808:Tetrahymena thermophila / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D019076:Transgenes; D014404:Tubulin / Q000235:genetics
D000953:Antigens, Protozoan; D000954:Antigens, Surface; C118810:IAG48 protein, Ichthyophthirius multifiliis; D008565:Membrane Proteins; D015800:Protozoan Proteins; D014404:Tubulin; D017239:Paclitaxel