02350nas a2200313   4500008004600000245014400046210006900190300001400259490000900273520122600282653003001508653001201538653002101550653002301571653003901594653003501633653004201668653003001710653003101740653003701771653003201808653002201840653005001862653002101912100001101933700002001944700002201964856005001986       Submitted                   eng d     00aRapid elevation of Inos and decreases in abundance of other proteins at pupal diapause termination in the flesh fly Sarcophaga crassipalpis0 aRapid elevation of Inos and decreases in abundance of other prot  a663 - 6680 v17943 aWe analyzed changes in brain proteins 24 h after pupal diapause termination in Sarcophaga crassipalpis by a combination of 2-dimensional gel electrophoresis and mass spectrometry. The proteome analysis revealed significant changes in 20 proteins, 11 of which represented &gt;or=2.5-fold changes. Three proteins were present only in the brains of diapausing pupae. Among the most abundant proteins that showed a change, 1 was more abundant, 7 were less abundant, and 2 were absent following diapause termination. The protein that increased in abundance following diapause termination showed highest identity to myo-inositol-1-phosphate synthase (Inos). Proteins that decreased at diapause termination included those showing highest identities to fatty acid binding protein, CG2331-PA, twinstar, catalase, and a histone. Proteins absent at diapause termination included ribosomal protein L17A and one unnamed protein. An increase of Inos protein level was confirmed using Western blot analysis. Attempts to terminate diapause by injection of several Inos-related metabolites failed, thus suggesting that the elevation of Inos at diapause termination is downstream of the physiological regulation that initiates development.10aAdaptation, Physiological10aAnimals10aBrain/metabolism10aDatabases, Protein10adevelopment/*metabolism/physiology10aDiptera/*metabolism/physiology10aElectrophoresis, Gel, Two-Dimensional10aGene Expression Profiling10aGene Expression Regulation10aInositol/metabolism/pharmacology10aInsect Proteins/*metabolism10aMass Spectrometry10aMyo-Inositol-1-Phosphate Synthase/*metabolism10aPupa/growth &amp1 aLi, A.1 aMichaud, M., R.1 aDenlinger, D., L.  uhttps://sarcophagidae.myspecies.info/node/28403073nas a2200289   4500008004600000245011700046210006900163300000800232490000700240520212900247653002902376653002002405653001202425653002202437653003002459653001702489653002702506653002602533653002102559653003602580653003602616100001702652700002002669700002202689700002202711856005002733       Submitted                   eng d     00aGene discovery using massively parallel pyrosequencing to develop ESTs for the flesh fly Sarcophaga crassipalpis0 aGene discovery using massively parallel pyrosequencing to develo  a2340 v103 aBACKGROUND: Flesh flies in the genus Sarcophaga are important models for investigating endocrinology, diapause, cold hardiness, reproduction, and immunity. Despite the prominence of Sarcophaga flesh flies as models for insect physiology and biochemistry, and in forensic studies, little genomic or transcriptomic data are available for members of this genus. We used massively parallel pyrosequencing on the Roche 454-FLX platform to produce a substantial EST dataset for the flesh fly Sarcophaga crassipalpis. To maximize sequence diversity, we pooled RNA extracted from whole bodies of all life stages and normalized the cDNA pool after reverse transcription. RESULTS: We obtained 207,110 ESTs with an average read length of 241 bp. These reads assembled into 20,995 contigs and 31,056 singletons. Using BLAST searches of the NR and NT databases we were able to identify 11,757 unique gene elements (E&lt;0.0001) representing approximately 9,000 independent transcripts. Comparison of the distribution of S. crassipalpis unigenes among GO Biological Process functional groups with that of the Drosophila melanogaster transcriptome suggests that our ESTs are broadly representative of the flesh fly transcriptome. Insertion and deletion errors in 454 sequencing present a serious hurdle to comparative transcriptome analysis. Aided by a new approach to correcting for these errors, we performed a comparative analysis of genetic divergence across GO categories among S. crassipalpis, D. melanogaster, and Anopheles gambiae. The results suggest that non-synonymous substitutions occur at similar rates across categories, although genes related to response to stimuli may evolve slightly faster. In addition, we identified over 500 potential microsatellite loci and more than 12,000 SNPs among our ESTs. CONCLUSION: Our data provides the first large-scale EST-project for flesh flies, a much-needed resource for exploring this model species. In addition, we identified a large number of potential microsatellite and SNP markers that could be used in population and systematic studies of S. crassipalpis and other flesh flies.10a*Expressed Sequence Tags10a*Genome, Insect10aAnimals10aDiptera/*genetics10aGene Expression Profiling10aGene Library10aMicrosatellite Repeats10aOID - NLM: PMC270081710aPMC - PMC270081710aPolymorphism, Single Nucleotide10aSequence Analysis, DNA/*methods1 aHahn, D., A.1 aRagland, G., J.1 aShoemaker, D., D.1 aDenlinger, D., L.  uhttps://sarcophagidae.myspecies.info/node/290