Four serine proteinase inhibitors (serpin) from the brown ear tick, Rhiphicephalus appendiculatus; cDNA cloning and preliminary characterization.
While development of an anti-Boophilus microplus vaccine is advanced and practical, work on other economically important ticks such as Rhipicephalus appendiculatus is still in its infancy. Guess PCR primers, designed from a consensus amino acid sequence (NAVYKFG) motif were used with rapid amplification of cDNA ends (RACE) to clone four cDNAs encoding serine proteinase inhibitors (serpin) from the brown ear tick Rhipicephalus appendiculatus. The four genes designated as R. appendiculatus serpin (RAS) -1 to -4 encode polypeptides of 378, 380, 398 and 486 amino acids long, respectively. Sequence comparison of RAS-1 to -4 predicted amino acid sequences to the serpin-like hypothetical protein from Ixodes ricinus (Leboulle et al., 2002) revealed closer structural similarities among tick serpins. Expression analysis by RT-PCR showed that RAS-1 to -4 are expressed in other tick organs in addition to salivary glands and midguts. Except for RAS-3 whose expression level appears to be equivalent in all tick organs, RAS-1, -2 and -4 are predominantly expressed in the salivary glands. We have discussed our findings with reference to development of vaccines against R. appendiculatus.
Mulenga
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A
Graduate School of Veterinary Medicine, Hokkaido University, Kitaku, Kita 18, Nishi 9, Sapporo 060-0818, Hokkaido, Japan. amvle001@umaryland.edu
Tsuda
A
A
Onuma
M
M
Sugimoto
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015842:Serine Proteinase Inhibitors / Q000235:genetics / Q000302:isolation & purification*; D015843:Serpins / Q000235:genetics / Q000302:isolation & purification*; D013987:Ticks / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D015843:Serpins