Surface display of a parasite antigen in the ciliate Tetrahymena thermophila

被引:66
作者
Gaertig, J [1 ]
Gao, Y
Tishgarten, T
Clark, TG
Dickerson, HW
机构
[1] Univ Georgia, Dept Cellular Biol, Athens, GA 30602 USA
[2] Univ Georgia, Dept Med Microbiol & Parasitol, Athens, GA 30602 USA
[3] Cornell Univ, Dept Microbiol & Immunol, Ithaca, NY 14853 USA
关键词
Tetrahymena; parasite antigen; beta-tubulin; paclitaxel; vaccine;
D O I
10.1038/8638
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 DMA, 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.
引用
收藏
页码:462 / 465
页数:4
相关论文
共 35 条
[1]  
Ausubel F.M., 1988, CURRENT PROTOCOLS MO
[2]  
CALZONE FJ, 1983, J BIOL CHEM, V258, P6887
[3]  
CALZONE FJ, 1982, THESIS U ROCHESTER R
[4]  
CassidyHanley D, 1997, GENETICS, V146, P135
[5]   DEVELOPMENTAL EXPRESSION OF SURFACE-ANTIGEN GENES IN THE PARASITIC CILIATE ICHTHYOPHTHIRIUS-MULTIFILIIS [J].
CLARK, TG ;
MCGRAW, RA ;
DICKERSON, HW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (14) :6363-6367
[6]   Surface antigen cross-linking triggers forced exit of a protozoan parasite from its host [J].
Clark, TG ;
Lin, TL ;
Dickerson, HW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (13) :6825-6829
[7]  
CLARK TG, 1996, ANN REV FISH DEV, V5, P113
[8]   ELECTROPORATION-MEDIATED REPLACEMENT OF A POSITIVELY AND NEGATIVELY SELECTABLE BETA-TUBULIN GENE IN TETRAHYMENA-THERMOPHILA [J].
GAERTIG, J ;
THATCHER, TH ;
GU, L ;
GOROVSKY, MA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (10) :4549-4553
[9]   HIGH-FREQUENCY VECTOR-MEDIATED TRANSFORMATION AND GENE REPLACEMENT IN TETRAHYMENA [J].
GAERTIG, J ;
GU, L ;
HAL, B ;
GOROVSKY, MA .
NUCLEIC ACIDS RESEARCH, 1994, 22 (24) :5391-5398
[10]   PERSPECTIVES ON TUBULIN ISOTYPE FUNCTION AND EVOLUTION BASED ON THE OBSERVATION THAT TETRAHYMENA-THERMOPHILA MICROTUBULES CONTAIN A SINGLE ALPHA-TUBULIN AND BETA-TUBULIN [J].
GAERTIG, J ;
THATCHER, TH ;
MCGRATH, KE ;
CALLAHAN, RC ;
GOROVSKY, MA .
CELL MOTILITY AND THE CYTOSKELETON, 1993, 25 (03) :243-253