Transcription Profiling

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Hilary Ranson - One of the best experts on this subject based on the ideXlab platform.

  • Transcription Profiling of a recently colonised pyrethroid resistant anopheles gambiae strain from ghana
    2007
    Co-Authors: Pie Muller, Martin J Donnelly, Hilary Ranson
    Abstract:

    Background Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases) have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown.

  • Transcription Profiling of a recently colonised pyrethroid resistant anopheles gambiae strain from ghana
    2007
    Co-Authors: Pie Muller, Martin J Donnelly, Hilary Ranson
    Abstract:

    Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases) have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown. Here, constitutive expression patterns of genes putatively involved in conferring pyrethroid resistance was investigated in a recently colonised pyrethroid resistant Anopheles gambiae strain from Odumasy, Southern Ghana. RNA from the resistant strain and a standard laboratory susceptible strain, of both sexes was extracted, reverse transcribed and labelled with either Cy3- or Cy5-dye. Labelled cDNA was co-hybridised to the detox chip, a custom-made microarray containing over 230 A. gambiae gene fragments predominantly from enzyme families associated with insecticide resistance. After hybridisation, Cy3- and Cy5-signal intensities were measured and compared gene by gene. In both females and males of the resistant strain the cytochrome P450s CYP6Z2 and CYP6M2 are highly over-expressed along with a member of the superoxide dismutase (SOD) gene family. These genes differ from those found up-regulated in East African strains of pyrethroid resistant A. gambiae and constitute a novel set of candidate genes implicated in insecticide detoxification. These data suggest that metabolic resistance may have multiple origins in A. gambiae, which has strong implications for the management of resistance.

Pie Muller - One of the best experts on this subject based on the ideXlab platform.

  • Transcription Profiling of a recently colonised pyrethroid resistant anopheles gambiae strain from ghana
    2007
    Co-Authors: Pie Muller, Martin J Donnelly, Hilary Ranson
    Abstract:

    Background Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases) have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown.

  • Transcription Profiling of a recently colonised pyrethroid resistant anopheles gambiae strain from ghana
    2007
    Co-Authors: Pie Muller, Martin J Donnelly, Hilary Ranson
    Abstract:

    Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases) have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown. Here, constitutive expression patterns of genes putatively involved in conferring pyrethroid resistance was investigated in a recently colonised pyrethroid resistant Anopheles gambiae strain from Odumasy, Southern Ghana. RNA from the resistant strain and a standard laboratory susceptible strain, of both sexes was extracted, reverse transcribed and labelled with either Cy3- or Cy5-dye. Labelled cDNA was co-hybridised to the detox chip, a custom-made microarray containing over 230 A. gambiae gene fragments predominantly from enzyme families associated with insecticide resistance. After hybridisation, Cy3- and Cy5-signal intensities were measured and compared gene by gene. In both females and males of the resistant strain the cytochrome P450s CYP6Z2 and CYP6M2 are highly over-expressed along with a member of the superoxide dismutase (SOD) gene family. These genes differ from those found up-regulated in East African strains of pyrethroid resistant A. gambiae and constitute a novel set of candidate genes implicated in insecticide detoxification. These data suggest that metabolic resistance may have multiple origins in A. gambiae, which has strong implications for the management of resistance.

Kenneth J Linthicum - One of the best experts on this subject based on the ideXlab platform.

  • Open Access Full Text Article
    2016
    Co-Authors: Daniel L Kline, Jian Chen, James J Becnel, Sandra A Allan, Gary G Clark, Kenneth J Linthicum, Correspondence Liming Zhao
    Abstract:

    Identification and Transcription Profiling of NDUFS8 in Aedes taeniorhynchus (Diptera: Culicidae): developmental regulation and environmental response Liming Zhao1–

  • identification and Transcription Profiling of ndufs8 in aedes taeniorhynchus diptera culicidae developmental regulation and environmental response
    2014
    Co-Authors: Liming Zhao, Jian Chen, Daniel L Kline, James J Becnel, Sandra A Allan, Gary G Clark, Kenneth J Linthicum
    Abstract:

    Abstract : The cDNA of a NADH dehydrogenase-ubiquinone Fe-S protein 8 subunit (NDUFS8) gene from Aedes (Ochlerotatus) taeniorhynchus Wiedemann has been cloned and sequenced. The 824 bp full-length mRNA sequence of AetNDUFS8 encodes an open reading region of 651 bp (that is, 217 amino acids). To identify if AetNDUFS8 is developmentally regulated, we employed a quantitative real-time polymerase chain reaction to examine AetNDUFS8 gene expression levels in all developmental stages of Ae. taeniorhynchus. In egg, larval, and pupal stages, AetNDUFS8 was expressed at relatively low levels. However, in the teneral adult stage, AetNDUFS8 was highly expressed. During the time course study in response to permethrin pesticide treatment, quantitative real-time polymerase chain reaction (PCR) also showed that mRNA Transcription levels of AetNDUFS8 were regulated in female Ae. taeniorhynchus. A mitochondrially encoded NADH dehydrogenase subunit 5 AetNADH5 was highly expressed in different developmental stages of Ae. taeniorhynchus. This study suggests that AetNDUFS8 and AetNADH5 play an essential role in the development of Ae. taeniorhynchus and will provide information useful for developing dsRNA pesticide for mosquito control.

  • identification and Transcription Profiling of trypsin in aedes taeniorhynchus diptera culicidae developmental regulation blood feeding and permethrin exposure
    2011
    Co-Authors: Liming Zhao, Jian Chen, Daniel L Kline, James J Becnel, Gary G Clark, Kenneth J Linthicum
    Abstract:

    The cDNA of a trypsin gene from Aedes (Ochlerotatus) taeniorhynchus (Weidemann) was cloned and sequenced. The full-length mRNA sequence (890 bp) for trypsin from Ae. taeniorhynchus (AetTryp1) was obtained, which encodes an open reading frame of 765 bp (i.e., 255 amino acids). To detect whether AetTryp is developmentally regulated, a quantitative real-time polymerase chain reaction was used to examine AetTrypl mRNA expression levels in different developmental stages of Ae. taeniorhynchus. AetTryp1 was expressed at low levels in egg, larval, and pupal stages, but was differentially expressed in adult Ae. taeniorhynchus, with highest levels found in 5-d-old female adults when compared with teneral adults. In addition, AetTryp1 mRNA expression differed between sexes, with expression levels much lower in males. However, in both males and females, there was a significant increase in AetTryp1 Transcription levels as age increased and peaked in 5-d-old adults. AetTrypl expressed in 5-d-old female Ae. taeniorhynchus significantly increased after 30 min postblood feeding compared with the control. The AetTryp1 mRNA expression in 5-d-old female Ae. taeniorhynchus was affected by different concentrations of permethrin.

Martin J Donnelly - One of the best experts on this subject based on the ideXlab platform.

  • Transcription Profiling of a recently colonised pyrethroid resistant anopheles gambiae strain from ghana
    2007
    Co-Authors: Pie Muller, Martin J Donnelly, Hilary Ranson
    Abstract:

    Background Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases) have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown.

  • Transcription Profiling of a recently colonised pyrethroid resistant anopheles gambiae strain from ghana
    2007
    Co-Authors: Pie Muller, Martin J Donnelly, Hilary Ranson
    Abstract:

    Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases) have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown. Here, constitutive expression patterns of genes putatively involved in conferring pyrethroid resistance was investigated in a recently colonised pyrethroid resistant Anopheles gambiae strain from Odumasy, Southern Ghana. RNA from the resistant strain and a standard laboratory susceptible strain, of both sexes was extracted, reverse transcribed and labelled with either Cy3- or Cy5-dye. Labelled cDNA was co-hybridised to the detox chip, a custom-made microarray containing over 230 A. gambiae gene fragments predominantly from enzyme families associated with insecticide resistance. After hybridisation, Cy3- and Cy5-signal intensities were measured and compared gene by gene. In both females and males of the resistant strain the cytochrome P450s CYP6Z2 and CYP6M2 are highly over-expressed along with a member of the superoxide dismutase (SOD) gene family. These genes differ from those found up-regulated in East African strains of pyrethroid resistant A. gambiae and constitute a novel set of candidate genes implicated in insecticide detoxification. These data suggest that metabolic resistance may have multiple origins in A. gambiae, which has strong implications for the management of resistance.

Donald A Bryant - One of the best experts on this subject based on the ideXlab platform.

  • Transcription Profiling of the model cyanobacterium synechococcus sp strain pcc 7002 by next gen solid sequencing of cdna
    2011
    Co-Authors: Marcus Ludwig, Donald A Bryant
    Abstract:

    The genome of the unicellular, euryhaline cyanobacterium Synechococcus sp. PCC 7002 encodes about 3200 proteins. Transcripts were detected for nearly all annotated open reading frames by a global transcriptomic analysis by Next-Generation (SOLiDTM) sequencing of cDNA. In the cDNA samples sequenced, ~90% of the mapped sequences were derived from the 16S and 23S ribosomal RNAs and ~10% of the sequences were derived from mRNAs. In cells grown photoautotrophically under standard conditions (38 °C, 1% (v/v) CO2 in air, 250 µmol photons m-2 s-1), the highest transcript levels (up to 2% of the total mRNA for the most abundantly transcribed genes (e. g., cpcAB, psbA, psaA)) were generally derived from genes encoding structural components of the photosynthetic apparatus. High light exposure for one hour caused changes in transcript levels for genes encoding proteins of the photosynthetic apparatus, Type-1 NADH dehydrogenase complex and ATP synthase, whereas dark incubation for one hour resulted in a global decrease in transcript levels for photosynthesis-related genes and an increase in transcript levels for genes involved in carbohydrate degradation. Transcript levels for pyruvate kinase and the pyruvate dehydrogenase complex decreased sharply in cells incubated in the dark. Under dark anoxic (fermentative) conditions, transcript changes indicated a global decrease in transcripts for respiratory proteins and suggested that cells employ an alternative phosphoenolpyruvate degradation pathway via phosphoenolpyruvate synthase (ppsA) and the pyruvate:ferredoxin oxidoreductase (nifJ). Finally, the data suggested that an apparent operon involved in tetrapyrrole biosynthesis and fatty acid desaturation, acsF2-ho2-hemN2-desF, may be regulated by oxygen concentration.

  • roles of xanthophyll carotenoids in protection against photoinhibition and oxidative stress in the cyanobacterium synechococcus sp strain pcc 7002
    2010
    Co-Authors: Joel E Graham, Marcus Ludwig, Wei Xiong, Richard M Alvey, Gaozhong Shen, Donald A Bryant
    Abstract:

    Abstract Synechococcus sp. strain PCC 7002 is a robust, genetically tractable cyanobacterium that produces six different xanthophyll carotenoids (zeaxanthin, cryptoxanthin, myxoxanthophyll (myxol-2′-fucoside), echinenone, 3′-hydroxyechinenone, and synechoxanthin) and tolerates many environmental stresses, including high light intensities. Targeted mutations were introduced to block the branches of the carotenoid biosynthetic pathway leading to specific xanthophylls, and a mutant lacking all xanthophylls was constructed. Some of the mutants showed severe growth defects at high light intensities, and multi-locus mutants had somewhat lower chlorophyll contents and lower photosystem I levels. The results suggested that xanthophylls, particularly zeaxanthin and echinenone, might play regulatory roles in thylakoid biogenesis. Measurements of reactive oxygen (ROS) and nitrogen (RNS) species in the mutants showed that all xanthophylls participate in preventing ROS/RNS accumulation and that a mutant lacking all xanthophylls accumulated very high levels of ROS/RNS. Results from Transcription Profiling showed that mRNA levels for most genes encoding the enzymes of carotenogenesis are significantly more abundant after exposure to high light. These studies indicated that all xanthophylls contribute to protection against photo-oxidative stress.