Cloning and disruption of the antigenic catalase gene of Aspergillus fumigatus

被引:103
作者
Calera, JA
Paris, S
Monod, M
Hamilton, AJ
Debeaupuis, JP
Diaquin, M
LopezMedrano, R
Leal, F
Latge, JP
机构
[1] INST PASTEUR, LAB ASPERGILLUS, F-75724 PARIS 15, FRANCE
[2] CHU VAUDOIS, SERV DERMATOL, CH-1011 LAUSANNE, SWITZERLAND
[3] GUYS HOSP, DERMATOL LAB, LONDON SE1 9RT, ENGLAND
[4] UNIV SALAMANCA, DEPT GENET & MICROBIOL, E-37008 SALAMANCA, SPAIN
关键词
D O I
10.1128/IAI.65.11.4718-4724.1997
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Aspergillus fumigatus possesses two catalases (described as fast and slow on the basis of their electrophoretic mobility). The slow catalase has been recognized as a diagnostic antigen for aspergillosis in immunocompetent patients. The antigenic catalase has been purified. The enzyme is a tetrameric protein composed of 90-kDa subunits. The corresponding cat1 gene was cloned, and sequencing data show that the cat1 gene codes for a 728-amino-acid polypeptide. A recombinant protein expressed in Pichia pastoris is enzymatically active and has biochemical and antigenic properties that are similar to those of the wild-type catalase. Molecular experiments reveal that CAT1 contains a signal peptide and a propeptide of 15 and 12 amino acrid residues, respectively. cat1-disrupted mutants that were unable to produce the slow catalase were as sensitive to H2O2 and polymorphonuclear cells as the wild-type strain. In addition, there was no difference in pathogenicity between the cat1 mutant and its parental cat1(+) strain in a murine model of aspergillosis.
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收藏
页码:4718 / 4724
页数:7
相关论文
共 49 条
[21]   Two divergent catalase genes are differentially regulated during Aspergillus nidulans development and oxidative stress [J].
Kawasaki, L ;
Wysong, D ;
Diamond, R ;
Aguirre, J .
JOURNAL OF BACTERIOLOGY, 1997, 179 (10) :3284-3292
[22]   PROPERTIES OF ASPERGILLUS-NIGER CATALASE [J].
KIKUCHITORII, K ;
HAYASHI, S ;
NAKAMOTO, H ;
NAKAMURA, S .
JOURNAL OF BIOCHEMISTRY, 1982, 92 (05) :1449-1456
[23]   IMMUNODIAGNOSIS OF ASPERGILLOSIS [J].
KURUP, VP ;
KUMAR, A .
CLINICAL MICROBIOLOGY REVIEWS, 1991, 4 (04) :439-456
[24]  
KY TV, 1968, REV IMMUNOL, V32, P37
[25]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+
[26]   A RAPID COLORIMETRIC ASSAY OF FUNGAL VIABILITY WITH THE TETRAZOLIUM SALT MTT [J].
LEVITZ, SM ;
DIAMOND, RD .
JOURNAL OF INFECTIOUS DISEASES, 1985, 152 (05) :938-945
[27]  
LEVITZ SM, 1985, J INFECT DIS, V152, P33, DOI 10.1093/infdis/152.1.33
[28]   Immunoblotting patterns in the serodiagnosis of aspergilloma: Antibody response to the 90 kDa Aspergillus fumigatus antigen [J].
LopezMedrano, R ;
Ovejero, MC ;
Calera, JA ;
Puente, P ;
Leal, F .
EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES, 1996, 15 (02) :146-152
[29]  
LOPEZMEDRANO R, 1995, INFECT IMMUN, V63, P4774
[30]  
LOPEZMEDRANO R, UNPUB