A CRYPTOSPORIDIUM-PARVUM GENOMIC REGION ENCODING HEMOLYTIC-ACTIVITY

被引:20
作者
STEELE, MI
KUHLS, TL
NIDA, K
MEKA, CSR
HALABI, IM
MOSIER, DA
ELLIOTT, W
CRAWFORD, DL
GREENFIELD, RA
机构
[1] UNIV OKLAHOMA,HLTH SCI CTR,DEPT INTERNAL MED,OKLAHOMA CITY,OK 73126
[2] KANSAS STATE UNIV AGR & APPL SCI,COLL VET MED,DEPT PATHOL,MANHATTAN,KS 66506
关键词
D O I
10.1128/IAI.63.10.3840-3845.1995
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Successful parasitization by Cryptosporidium parvum requires multiple disruptions in both host and protozoan cell membranes as cryptosporidial sporozoites invade intestinal epithelial cells and subsequently develop into asexual and sexual life stages. To identify cryptosporidial proteins which may play a role in these membrane alterations, hemolytic activity was used as a marker to screen a C. parvum genomic expression library. A stable hemolytic clone (H4) containing a 5.5-kb cryptosporidial genomic fragment was identified. The hemolytic activity encoded on H4 was mapped to a 1-kb region that contained a complete 690-bp open reading frame (hemA) ending in a common stop codon. A 21-kDa plasmid-encoded recombinant protein was expressed in maxicells containing H4. Subclones of H4 which contained only a portion of hemA did not induce hemolysis on blood agar or promote expression of the recombinant protein in maxicells. Reverse transcriptase-mediated PCR analysis of total RNA isolated from excysted sporozoites and the intestines of infected adult mice with severe combined immunodeficiency demonstrated that hemA is actively transcribed during the cryptosporidial life cycle.
引用
收藏
页码:3840 / 3845
页数:6
相关论文
共 42 条
[1]   PBLUESCRIPT-II - GENE-MAPPING VECTORS [J].
ALTINGMEES, MA ;
SHORT, JM .
NUCLEIC ACIDS RESEARCH, 1989, 17 (22) :9494-9494
[2]   BASIC LOCAL ALIGNMENT SEARCH TOOL [J].
ALTSCHUL, SF ;
GISH, W ;
MILLER, W ;
MYERS, EW ;
LIPMAN, DJ .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) :403-410
[3]   A TRYPANOSOMA-CRUZI SECRETED PROTEIN IMMUNOLOGICALLY RELATED TO THE COMPLEMENT COMPONENT-C9 - EVIDENCE FOR MEMBRANE PORE-FORMING ACTIVITY AT LOW PH [J].
ANDREWS, NW ;
ABRAMS, CK ;
SLATIN, SL ;
GRIFFITHS, G .
CELL, 1990, 61 (07) :1277-1287
[4]   THE ACID-ACTIVE HEMOLYSIN OF TRYPANOSOMA-CRUZI [J].
ANDREWS, NW .
EXPERIMENTAL PARASITOLOGY, 1990, 71 (02) :241-244
[5]   EMERGING INFECTIOUS-DISEASES IN THE UNITED-STATES, 1993 [J].
BERKELMAN, RL .
JOURNAL OF INFECTIOUS DISEASES, 1994, 170 (02) :272-277
[6]   CLONING OF A CDNA-ENCODING THE DENSE GRANULE PROTEIN GRA3 FROM TOXOPLASMA-GONDII [J].
BERMUDES, D ;
DUBREMETZ, JF ;
ACHBAROU, A ;
JOINER, KA .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1994, 68 (02) :247-257
[7]  
BREATHNACH R, 1981, ANNU REV BIOCHEM, V50, P349, DOI 10.1146/annurev.bi.50.070181.002025
[8]   REGULATION OF CYTOKINE PRODUCTION DURING THE EXPRESSION PHASE OF THE ANTICRYPTOCOCCAL DELAYED-TYPE HYPERSENSITIVITY RESPONSE [J].
BUCHANAN, KL ;
MURPHY, JW .
INFECTION AND IMMUNITY, 1994, 62 (07) :2930-2939
[9]   CRYPTOSPORIDIOSIS [J].
CURRENT, WL ;
GARCIA, LS .
CLINICAL MICROBIOLOGY REVIEWS, 1991, 4 (03) :325-358
[10]   A COMPREHENSIVE SET OF SEQUENCE-ANALYSIS PROGRAMS FOR THE VAX [J].
DEVEREUX, J ;
HAEBERLI, P ;
SMITHIES, O .
NUCLEIC ACIDS RESEARCH, 1984, 12 (01) :387-395