RESUMO
We created a cDNA library from feeding, female Ixodes scapularis ticks and screened the library with a subtracted probe to eliminate most genes common to feeding female and mating male I. scapularis ticks. Four unique genes were identified in this screen. One gene, Is 9, (represented by 16 cDNAs) was more highly expressed in female ticks. This gene encodes a putative glycine-rich protein, which matched a number of glycine-rich proteins including attachment cement proteins from Rhipicephalus appendiculatus. A second gene, Is 10 (represented by one cDNA) was also more highly expressed in female ticks, but did not match any other sequences in the GenBank database. The third gene, Is 11 (represented by one cDNA) was very similar to Drosophila sp. hsp68 and hsp70 genes and was expressed about equally in male and female ticks. The fourth gene, Is 12 (represented by two cDNAs) was also about equally expressed in male and female ticks, and was similar to a salivary gland gene from Ixodes ricinis. This gene also showed limited similarity to some cuticle genes from insects.
Assuntos
Ixodes/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Cervos/parasitologia , Comportamento Alimentar , Feminino , Regulação da Expressão Gênica/fisiologia , Biblioteca Gênica , Ixodes/metabolismo , Masculino , Dados de Sequência Molecular , RNA/química , RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
We tested 103 adult Ixodes scapularis Say from 12 counties in Minnesota for the presence of Borrelia burgdorferi and the causative agent of human granulocytic ehrlichiosis (HGE), using polymerase chain reaction (PCR). A total of 17 ticks (16.5%) was positive for B. burgdoiferi using nested PCR for the flagellin gene. or both PCR for the ospA gene and nested PCR for the flagellin gene. A total of four ticks (3.8%) was positive for the agent of HGE using nested PCR for 16S rDNA. Counties in Minnesota with established and recently reported populations of I. scapularis both had ticks infected with B. burgdorferi. The agent of HGE was only detected in counties with established I. scapularis populations.
Assuntos
Anaplasmataceae/isolamento & purificação , Borrelia burgdorferi/isolamento & purificação , Ixodes/microbiologia , Lipoproteínas , Infestações por Carrapato/parasitologia , Anaplasmataceae/genética , Animais , Antígenos de Superfície/genética , Proteínas da Membrana Bacteriana Externa/genética , Vacinas Bacterianas , Borrelia burgdorferi/genética , Doenças do Gato/parasitologia , Gatos , DNA/análise , DNA Bacteriano/análise , Cervos , Doenças do Cão/parasitologia , Cães , Ehrlichiose/microbiologia , Feminino , Flagelina/genética , Granulócitos , Doenças dos Cavalos/parasitologia , Cavalos , Humanos , Minnesota , Reação em Cadeia da Polimerase/métodos , Infestações por Carrapato/veterináriaRESUMO
We created a cDNA library from mating, male Ixodes scapularis ticks and screened the library with a subtracted probe to eliminate genes common to feeding female and mating male I. scapularis ticks. A total of seven unique cDNAs were identified in this screen. One cDNA had sequence similarity to the IGBP-MC gene from Rhipicephalus appendiculatus, and another cDNA potentially encodes a protein with similarity to metalloproteases. RT-PCR, using RNA isolated from male and female I. scapularis ticks, confirmed that these genes are expressed in male, but not female ticks. The remaining five cDNAs did not match any sequences in the GenBank database.