Project description:Babesia infected (and control) ovary tissue was used to construct LongSAGE libraries. The intent was to find differential gene expression in both libraries. Details: The extraction was done using Ambion's ToTALLY RNA kit, and the concatemers where constructed using Invitrogen's I-SAGE Long kit. The concatmer files (multiple fasta sequence files) were processed using in-house perl scripts to extract the 17bp LongSAGE tags. **Note: contact person: Felix D. Guerrero. email: felix.guerrero@ars.usda.gov Keywords: LongSAGE
Project description:Babesia bovis exposed tissues: Three tissues were looked at. 1. Adult Female Gut 2. Adult Ovary 3. Larvae. In all there are 24 measurements for feature (EST), and 4 measurements per treatment for each of the 6 treatment groups. The 6 treatment groups are: Gut infected (GI), Gut control (GC), and similarly for Ovary and Larval: OI, OC, LI, LC. There are 12 chips, each with a spot replicate. **Note: contact person: Felix D. Guerrero email: felix.guerrero@ars.usda.gov Keywords: Rhipicephalus (Boophilus) microplus, Babesia, microarrays.
Project description:Babesia bovis exposed tissues: Three tissues were looked at. 1. Adult Female Gut 2. Adult Ovary 3. Larvae. In all there are 24 measurements for feature (EST), and 4 measurements per treatment for each of the 6 treatment groups. The 6 treatment groups are: Gut infected (GI), Gut control (GC), and similarly for Ovary and Larval: OI, OC, LI, LC. There are 12 chips, each with a spot replicate. **Note: contact person: Felix D. Guerrero email: felix.guerrero@ars.usda.gov Keywords: Rhipicephalus (Boophilus) microplus, Babesia, microarrays. In the study presented here, 12 hybridizations were performed. Expression profiles for 13601 unique ESTs were analyzed. Uninfected ticks were collected in one batch, as were the infected ticks. Replicates performed are technical.
Project description:To understand the effect of Babesia infection on Rhipicephalus microplus hemocyte gene expression, we performed high throughput RNA-sequencing using samples collected from Rhipicephalus microplus uninfected tick hemolymph and infected with either Babesia bigemina or Babesia bovis. We evaluated gene expression differences that may be attributed to tick immune defense to babesial infection.
Project description:A R. microplus microarray was used to study differential gene expression in acaricide exposed larvae from an amitraz-resistant strain. The acaricide treatments were: organophosphate (OP), pyrethroid, ivermectin, and amitraz. The microarrays contained over 13,000 probes corresponding to each member of R. microplus gene index ESTs previously described (http://compbio.dfci.harvard.edu/tgi/cgi-bin/tgi/gimain.pl?gudb=b_microplus). Serial analysis of gene expression (SAGE) data from the OP treated R. microplus was used to verify the OP microarray data. The expression profiles of selected transcripts were verified by real time PCR. Among the significantly differentially expressed genes, were a tick legumain, involved in blood digestion, gluthathione S-transferase (GST), a detoxification enzyme involved in pesticide resistance, acyltransferase, several putative salivary sulfotransferases, and a glutamate receptor. **Note: contact person: Felix D. Guerrero email: felix.guerrero@ars.usda.gov Keywords: Rhipicephalus (Boophilus) microplus, acaricide resistance genes, organophosphates OP, microarrays, detoxification enzymes.
Project description:Transcriptional profiling of salivary gland, midgut and ovary tissues isolated from Rhipicephalus microplus females at day 20 post infestation. This enabled the identification of transcripts that are tissue-specific or shared among the tissues tested.
Project description:A R. microplus microarray was used to study differential gene expression in acaricide exposed larvae from an amitraz-resistant strain. The acaricide treatments were: organophosphate (OP), pyrethroid, ivermectin, and amitraz. The microarrays contained over 13,000 probes corresponding to each member of R. microplus gene index ESTs previously described (http://compbio.dfci.harvard.edu/tgi/cgi-bin/tgi/gimain.pl?gudb=b_microplus). Serial analysis of gene expression (SAGE) data from the OP treated R. microplus was used to verify the OP microarray data. The expression profiles of selected transcripts were verified by real time PCR. Among the significantly differentially expressed genes, were a tick legumain, involved in blood digestion, gluthathione S-transferase (GST), a detoxification enzyme involved in pesticide resistance, acyltransferase, several putative salivary sulfotransferases, and a glutamate receptor. **Note: contact person: Felix D. Guerrero email: felix.guerrero@ars.usda.gov Keywords: Rhipicephalus (Boophilus) microplus, acaricide resistance genes, organophosphates OP, microarrays, detoxification enzymes. In the study presented here, 10 hybridizations were preformed. Two technical replicates for each treatment. There were no biological replicates included in this study due to limitations on biomaterials. Expression profiles for 13601 unique ESTs were analyzed.
Project description:Organophosphate resistant and susceptible tick larvae from laboratory strains of the southern cattle tick, Rhipicephalus (Boophilus) microplus were exposed to low doses of the organophosphate (OP) acaricide, coumaphos. Serial Analysis of Gene Expression (SAGE) was used to analyze differential gene expression in response to OP treatment and to compare the responses in OP-treated and untreated resistant and susceptible tick larvae. ** note: Contact person is Felix D. Guerrero. email: felix.guerrero@ars.usda.gov Keywords: SAGE, acaricide response, organophosphate.
