Surface display of a parasite antigen in the ciliate Tetrahymena thermophila

Nat Biotechnol. 1999 May;17(5):462-5. doi: 10.1038/8638.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antigens, Protozoan / genetics*
  • Antigens, Protozoan / metabolism
  • Antigens, Surface / genetics
  • Antigens, Surface / metabolism
  • Ciliophora / genetics*
  • Ciliophora / metabolism
  • Cloning, Molecular / methods*
  • Drug Resistance / genetics
  • Fish Diseases / parasitology
  • Gene Expression
  • Immunoblotting
  • Immunohistochemistry
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Paclitaxel / pharmacology
  • Protozoan Proteins*
  • Recombination, Genetic
  • Tetrahymena thermophila / drug effects
  • Tetrahymena thermophila / genetics*
  • Tetrahymena thermophila / metabolism
  • Transgenes
  • Tubulin / genetics

Substances

  • Antigens, Protozoan
  • Antigens, Surface
  • IAG48 protein, Ichthyophthirius multifiliis
  • Membrane Proteins
  • Protozoan Proteins
  • Tubulin
  • Paclitaxel