Mining an Ostrinia nubilalis midgut expressed sequence tag (EST) library for candidate genes and single nucleotide polymorphisms (SNPs)

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2008-12-01
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Coates, Brad
Sumerford, Douglas
Lewis, Leslie
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Hellmich, Richard
Emeritus USDA-ARS Research Entomologist Emeritus Affiliate Professor
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Entomology

The Department of Entomology seeks to teach the study of insects, their life-cycles, and the practicalities in dealing with them, for use in the fields of business, industry, education, and public health. The study of entomology can be applied towards evolution and ecological sciences, and insects’ relationships with other organisms & humans, or towards an agricultural or horticultural focus, focusing more on pest-control and management.

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The Department of Entomology was founded in 1975 as a result of the division of the Department of Zoology and Entomology.

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Genes expressed in lepidopteran midgut tissues are involved in digestion and Bacillus thuringiensis (Bt) toxin resistance traits. Five hundred and thirty five unique transcripts were annotated from 1745 high quality O. nubilalis larval midgut expressed sequence tags (ESTs). Full-length cDNA sequence of 12 putative serine proteinase genes and 3 partial O. nubilalis aminopeptidase N protein genes, apn1, apn3, and apn4, were obtained, and genes may have roles in plant feeding and Bt toxin resistance traits of Ostrinia larvae. The EST library was not normalized and insert frequencies reflect transcript levels under the initial treatment conditions and redundancy of inserts from highly expressed transcripts allowed prediction of putative single nucleotide polymorphisms (SNPs). Ten di-, tri- or tetranucleotide repeat unit microsatellite loci were identified, and minisatellite repeats were observed within the C-termini of two encoded serine proteinases. Molecular markers showed polymorphism at 28 SNP loci and one microsatellite locus, and Mendelian inheritance indicated that markers were applicable to genome mapping applications. This O. nubilalis larval midgut EST collection is a resource for gene discovery, expression information, and allelic variation for use in genetic marker development.

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This article is from Insect Molecular Biology; 17 (2008); 607-620; doi: 10.1111/j.1365-2583.2008.00833.x

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