Characterisation of the gene encoding a 104-kilodalton microneme-rhoptry protein of Theileria parva.

Abstract:

:A neutralizing antiserum, C16, raised against sporozoites of Theileria parva parva was used to screen a lambda gt11 expression library of T. parva parva (Muguga) genomic DNA fragments. Proteins encoded by one phage clone, lambda TpS-17, were reactive with the C16 antiserum. Detailed characterisation of the DNA insert showed it to encode determinants found on four theilerial antigens of approximately 104, 90, 85 and 35 kDa. The sequence encoded by the clone is expressed during sporogony as a single RNA transcript of about 3000 nucleotides. On sequencing a portion of the 5000-bp insert, an open reading frame of 2772 bp was revealed that encoded a 104-kDa protein. Immunoscreening a library of subfragments of the DNA insert with the original antiserum localised sequences encoding the dominant antigenic determinants to an 800-bp stretch of DNA at the 3' end of the open reading frame. Sequence data from three subclones spanning this region show portions of the antigenic domains to be unusually rich in proline residues which are repeated every three amino acids. These repeats often take the form X-S(T)-P or X-K(R)-P. Antibodies directed against each of the three subclones recognize the 104- and 35-kDa antigens and different combinations of the 90- and 85-kDa kDa antigens, suggesting that the smaller proteins are derived from the 104-kDa antigen by limited proteolysis occurring at the carboxyl terminus end of the protein. In immunoelectron micrographs the antigen is associated with the microneme/rhoptry complexes of the sporozoite.

journal_name

Mol Biochem Parasitol

authors

Iams KP,Young JR,Nene V,Desai J,Webster P,ole-MoiYoi OK,Musoke AJ

doi

10.1016/0166-6851(90)90007-9

subject

Has Abstract

pub_date

1990-02-01 00:00:00

pages

47-60

issue

1

eissn

0166-6851

issn

1872-9428

pii

0166-6851(90)90007-9

journal_volume

39

pub_type

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