Characterization of a surface antigen of Eimeria tenella sporozoites and synthesis from a cloned cDNA in Escherichia coli.
An antigenic surface protein of Eimeria tenella sporozoites has been identified that is the target of two neutralizing monoclonal antibodies Ptn 7.2A4/4 and Ptn 9.9D12. The antigen as isolated from the parasite is composed of a 17 kDa polypeptide and a 8 kDa polypeptide linked by a disulfide bridge. De novo synthesis of the antigen does not begin until approximately 16-20 h after the initiation of oocyst sporulation. A cDNA library was constructed using mRNA from sporulated oocysts and a clone encoding the antigen was isolated. The Ta4 gene encodes a single polypeptide of 25 kDa which contains the 17 and 8 kDa polypeptides. The protein has been synthesized in Escherichia coli either directly or as part of a beta-galactosidase fusion protein. The products synthesized in E. coli are single polypeptides and are not cleaved to two polypeptides as is seen in the parasite. The products accumulate in bacteria in an insoluble form which can be solubilized and renatured to an immunoreactive form.
Brothers
V M
VM
Codon, South San Francisco, CA 94080.
Kuhn
I
I
Paul
L S
LS
Gabe
J D
JD
Andrews
W H
WH
Sias
S R
SR
McCaman
M T
MT
Dragon
E A
EA
Files
J G
JG
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D000954:Antigens, Surface / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004539:Eimeria / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D007118:Immunoassay; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000276:immunology; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D000953:Antigens, Protozoan; D000954:Antigens, Surface; D010455:Peptides; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D004247:DNA