Cloning and expression of a protective antigen from the cattle tick Boophilus microplus.
Glycoproteins located on the luminal surface of the plasma membrane of tick gut epithelial cells, when used to vaccinate cattle, are capable of stimulating an immune response that protects cattle against subsequent tick infestation. One such tick gut glycoprotein, designated Bm86, has been purified to homogeneity and the amino acid sequences of peptide fragments generated by endoproteinase Lys-C digestion have been determined. We report here the isolation and characterization of a cDNA that encodes Bm86. The nucleotide sequence of the cDNA contains a 1982-base-pair open reading frame and predicts that Bm86 contains 650 amino acids including a 19-amino acid signal sequence and a 23-amino acid hydrophobic region adjacent to the carboxyl terminus. The main feature of the deduced protein sequence is the repeated pattern of 6 cysteine residues, suggesting the presence of several epidermal growth factor-like domains. A fusion protein consisting of 599 amino acids of Bm86 and 651 amino acids of beta-galactosidase was expressed in Escherichia coli as inclusion bodies. Ticks engorging on cattle vaccinated with these inclusion bodies were significantly damaged as a result of the immune response against the cloned antigen.
Rand
K N
KN
Biotechnology Australia Pty. Ltd., Roseville, New South Wales.
Moore
T
T
Sriskantha
A
A
Spring
K
K
Tellam
R
R
Willadsen
P
P
Cobon
G S
GS
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D002417:Cattle / Q000469:parasitology; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005796:Genes; D005822:Genetic Vectors; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid; D013987:Ticks / Q000235:genetics* / Q000276:immunology; D014611:Vaccination
D000941:Antigens; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D004247:DNA