Molecular characterization of protease inhibitors from the Hessian fly, [Mayetiola destructor (Say)]

dc.contributor.authorMaddur, Appajaiah Ashoka
dc.date.accessioned2005-11-30T16:28:14Z
dc.date.available2005-11-30T16:28:14Z
dc.date.graduationmonthDecember
dc.date.issued2005-11-30T16:28:14Z
dc.date.published2005
dc.description.abstractAnalysis of transcriptomes from salivary glands and midgut of the Hessian fly [Mayetiola destructor (Say)] identified a diverse set of cDNAs that were categorized into five groups, group I – V, based on their phylogenetic relationship. All five of these groups may encode putative protease inhibitors based on structural similarity with known proteins. The sequences of these putative proteins among different groups are highly diversified. However, sequence identity and structural analysis of the proteins revealed that all of them contained high cysteine residues that were completely conserved at their respective positions among these otherwise diversified proteins. Analysis of bacterial artificial chromosome (BAC) DNA for two groups, group I (11A6) and group II (14A4), indicated that group I might be a single copy gene or genes with low copy number whereas group II exists as multiple copies clustered within the Hessian fly genome. To test the inhibitory activity and specificity of these putative proteins, recombinant proteins were generated. Enzymatic analysis of the recombinant proteins against commercial and insect gut proteases demonstrated that recombinant proteins indeed are strong inhibitors of proteases with different specificities. Northern analysis of the representative members of five groups revealed that the group I-IV genes were expressed exclusively in the larval stage with variations among groups at different larval stages. The group V (11C4) genes were expressed in the late larval and pupal stage. Tissue specific gene expression analysis revealed that group I-IV genes were predominantly expressed in malpighian tubules whereas the group V genes were abundantly expressed in the salivary glands. Localization experiments with the antibody for representative members from group II (14A4) demonstrated that the protein was predominantly localized in the malpighian tubules and in low amounts in the midgut, suggesting that malpighian tubules are the primary tissue of 14A4 inhibitor synthesis. The overall results indicated that the Hessian fly contains a complex network of genes that code for protease inhibitors which regulate protease activities through different developmental stages of the insect.
dc.description.advisorMing-Shun Chen
dc.description.advisorGerald E. Wilde
dc.description.degreeDoctor of Philosophy
dc.description.departmentDepartment of Entomology
dc.description.levelDoctoral
dc.format.extent1758436 bytes
dc.format.mimetypeapplication/pdf
dc.identifier.urihttp://hdl.handle.net/2097/136
dc.language.isoen_US
dc.publisherKansas State University
dc.rights© the author. This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/
dc.subjectProtease inhibitors
dc.subjectProteolysis
dc.subjectcDNA library
dc.subjectProteases
dc.subjectInhibition
dc.subjectGene expression
dc.subjectSalivary glands
dc.subjectHessian fly
dc.subject.umiBiology, Entomology (0353)
dc.subject.umiBiology, Genetics (0369)
dc.subject.umiBiology, Molecular (0307)
dc.titleMolecular characterization of protease inhibitors from the Hessian fly, [Mayetiola destructor (Say)]
dc.typeDissertation

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