Cloning and partial characterization of a Boophilus microplus (Acari: Ixodidae) calreticulin.
We report the cloning, sequence characterization and expression analysis of a calreticulin (CRT) coding cDNA of Boophilus microplus. CRT is a calcium-binding protein involved in multiple cell functions and possibly implicated in parasites host immune system evasion. The CRT cDNA sequence and its molecular characterization are described. Sequence similarity and phylogenetic analyses indicate a close relationship to other arthropod CRT sequences. The CRT cDNA was also expressed in a procariotic system and the recombinant protein (rBmCRT) was used to raise antibodies in a rabbit. Expression analyses of the corresponding gene in different developmental stages and tissues were performed by RT-PCR and Western-blot, which indicated a ubiquitous expression of the B. microplus calreticulin gene and demonstrated its presence in saliva. Sera of tick-infested bovines suggested that this protein may not be able to induce an IgG-based humoral response in its natural host.
Ferreira
Carlos Alexandre Sanchez
CA
Centro de Biotecnologia do Estado do Rio Grande do Sul, Universidade Federal do Rio Grande do Sul, Caixa Postal 15005, Campus do Vale, Porto Alegre, RS, Brazil.
Da Silva Vaz
Itabajara
I
da Silva
Sérgio Silva
SS
Haag
Karen L
KL
Valenzuela
Jesus G
JG
Masuda
Aoi
A
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D001483:Base Sequence; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D037282:Calreticulin; D002417:Cattle; D002418:Cattle Diseases / Q000276:immunology / Q000469:parasitology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D005260:Female; D005786:Gene Expression Regulation; D026863:Ixodidae / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D013984:Tick Infestations / Q000276:immunology / Q000662:veterinary
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D018076:DNA, Complementary; D012261:Ribonucleoproteins