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Poster 5:Identification of 5-Enolpyruvylshikimate-3-Phosphate Synthase in Various Grass SpeciesEric J. Gustafson / Faculty Sponsor: Dr. David RobinsonThe objective of this experiment was to explore the possibility of glyphosate resistanceoccurring within six various species of grasses by cloning the gene forherbicide resistance and treating plants with glyphosate. The gene of interest was5-Enolpyruvylshikimate-3-Phosphate Synthase (EPSPS), which becomes mutatedcausing this gene to activate, thus causing herbicide resistance to glyphosate, theactive ingredient in Roundup. Six grass species: Alliance Rye (Dactylis glomerata),Wolfpack Tall Fescue (Euphorbia pulcherrima), Annual Rye (Lolium multiflorum),Creeping Red Fescue (Festuca rubra), Annual Bluegrass (Poa pratensis), and A1A4Bentgrass (Agrostis palustris), were used in this study. Primers were used to amplifythe EPSPS gene in each species and gels were run to check for presence, visualizationyielded positive results for the presence in each species. For cloning, an optimizationPCR was run to extract a DNA sample via gel extraction. PCR product wasthen cloned and plated onto LB agar plates containing Ampicillin, and incubated at37°C overnight. Colonies were plucked to subculture using LB broth with an antibiotic,and plasmids were purified. Purified plasmid products were then digested withan EcoRI digest and the bands were observed by gel electrophoresis and a samplewas chosen with a potential clone. DNA quantification of the plasmid was runand the sample was diluted for sequencing at U of L Sequencing Core to comparethrough BLAST searches. Further plants will be tested for potential cloning andsequence comparing in the future.Poster 6:Further Characterization of a Ragweed PollencDNA LibraryMcKenna Ennes / Faculty Sponsor: Dr. David RobinsonIt is estimated that approximately 10 percent of all Americans are allergic to thepollen produced by the plant commonly known as Giant Ragweed (Ambrosiatrifida L.). Bellarmine University has created a cDNA library of pollen from thisspecies. So far only 134 sequences have been published in the NCBI GenBank. Theobjectives of this research were to sequence 96 more genes and identify them andpredict their function, metabolic role, and cellular location. Individual clones weregrown and plasmid was isolated, quantified, and digested with restriction enzyme topredict size. Sizes of the cDNA clones varied from approximately 250bp to 1500bp.Following sequencing, cDNA was examined using the Basic Local AlignmentSearch Tool (BLAST) to potentially identify the function of the gene product. Thisinformation will be useful in describing the biological and biochemical make up ofthis important health issue.6
Poster 7:Continued Studies of Ventral Eversible Gland Functionand Composition in Velvetbean Caterpillars,Anticarsia gemmatalis (Lepidoptera: Noctuidae)Elizabeth A. Shaw / Faculty Sponsor: Dr. Anthony LentzThe function and composition of the ventral eversible gland of Anticarsia gemmataliscaterpillars (Lepidoptera: Noctuidae) was studied. Larvae were reared in thelaboratory and fed a synthetic diet. The ventral eversible gland was removed fromhalf of the A. gemmatalis specimens and the remaining larvae were subjected to asham dissection procedure, but the gland was not removed. All larvae were allowedto feed as before and were weighed each day until pupation. Daily observations,including date of pupation and eclosion were recorded. Glands were also collectedand extracted in dichloromethane to analyze volatile content by gas chromatographycoupled with mass spectrometry. No differences were found between (a) the weightsof gland-dissected and control larvae or (b) pupation cycles, thus no conclusionscould be drawn about the gland’s function in relation to larval development. Resultsfrom GC/MS analysis suggest several possible organic compounds in the ventraleversible gland and/or gland secretions.Poster 8:Do single nucleotide polymorphisms affectthe synthesis of Brain-Derived NeurotrophicFactor (BDNF) in lymphoblast cells frombipolar disorder patients?Mathew Galante / Faculty Sponsor: Dr. David RobinsonAlterations in the expression of brain-derived neurotrophic factor (BDNF) may beinvolved in the development of bipolar disorder. BDNF is a neurotrophin that actsas a signaling molecule responsible for dendritic growth, neuronal plasticity, andlong-term potentiation. It also plays an important role in promoting the development,regeneration, survival, and maintenance of neuronal activity. The BDNFprotein is synthesized as a precursor, proBDNF, which is cleaved to the mature14-kDa form, mBDNF. Through interactions with the p75 NTR receptor, proBDNFelicits cellular apoptosis, while mBDNF, through the TrkB receptor, causes cellularproliferation. Previous genetic studies have shown single nucleotide polymorphisms(SNPs) within the BDNF gene may increase the risk of developing bipolar disorder.Studies have indicated abnormalities in ionic regulations play an important rolein the pathophysiology of bipolar disorder. The aim of this study was to examineintracellular BDNF expression and the functional effect of BDNF SNPs on ionicstress-induced apoptosis in bipolar disorder patients. Through a double-blindexperiment thirty cell lines from bipolar disorder patients were tested to determinewhether mutated BDNF altered expression, and how elevated intracellular sodiumconcentrations caused cellular apoptosis. It was determined that neither proBDNFnor mBDNF protein levels varied with the presence of SNPs believed to be associatedwith the risk for bipolar disorder.7
Page 1 and 2: BELLARMINE UNIVERSITYPresentsthe Ni
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