Document Detail


Cloning and expression of a DNA sequence encoding a 41-kilodalton Cryptosporidium parvum oocyst wall protein.
MedLine Citation:
PMID:  10548585     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This study was conducted to produce a recombinant species-specific oocyst wall protein of Cryptosporidium parvum. Antigens unique to C. parvum were identified by gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of oocyst proteins from several different Cryptosporidium species. Antiserum was then prepared against a 41-kDa antigen unique to C. parvum and used to identify a recombinant DNA clone, designated rCP41. Expression of CP41 mRNA in C. parvum oocysts was confirmed by reverse transcriptase PCR (RT-PCR). Although the CP41 sequence was shown by PCR to be present in the genome of C. baileyi, CP41 mRNA was not detected in this species by RT-PCR. Immunofluorescence staining with antiserum against recombinant CP41 detected native CP41 antigen on the surface of C. parvum oocysts but failed to detect CP41 on C. baileyi oocysts. Immunoelectron microscopy demonstrated that native CP41 was distributed unevenly on the C. parvum oocyst surface and was associated with amorphous oocyst wall material. In an enzyme-linked immunosorbent assay, purified rCP41 performed as well as native C. parvum oocyst protein in measuring the serological responses of young calves and adult cows to experimental and natural C. parvum infections. These results indicate that recombinant CP41 antigen may have potential in the immunodiagnosis of cryptosporidiosis.
Authors:
M C Jenkins; J Trout; C Murphy; J A Harp; J Higgins; W Wergin; R Fayer
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Clinical and diagnostic laboratory immunology     Volume:  6     ISSN:  1071-412X     ISO Abbreviation:  Clin. Diagn. Lab. Immunol.     Publication Date:  1999 Nov 
Date Detail:
Created Date:  2000-03-08     Completed Date:  2000-03-08     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  9421292     Medline TA:  Clin Diagn Lab Immunol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  912-20     Citation Subset:  IM    
Affiliation:
Immunology and Disease Resistance Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland 20705, USA. mjenkins@lpsi.barc.usda.gov
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AF144621
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Animals
Antibodies, Protozoan
Antigens, Protozoan / genetics*,  immunology
Base Sequence
Blotting, Northern
Cattle
Cell Wall / chemistry,  ultrastructure
Cloning, Molecular
Cryptosporidiosis / diagnosis*
Cryptosporidium parvum / genetics*,  growth & development,  immunology
DNA Primers
DNA, Protozoan / analysis
Enzyme-Linked Immunosorbent Assay
Gene Expression
Humans
Microscopy, Immunoelectron
Molecular Sequence Data
Protozoan Proteins / genetics*,  immunology
RNA, Messenger / analysis
Rabbits
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, DNA
Species Specificity
Chemical
Reg. No./Substance:
0/Antibodies, Protozoan; 0/Antigens, Protozoan; 0/CP41 protein, Cryptosporidium; 0/DNA Primers; 0/DNA, Protozoan; 0/Protozoan Proteins; 0/RNA, Messenger
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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