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Construction and Characterization of an Expressed Sequenced Tag Library for the Mosquito Vector Armigeres Subalbatus George F

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Entomology Publications Entomology 2007 Construction and characterization of an expressed sequenced tag library for the mosquito vector Armigeres subalbatus George F. Mayhew University of Wisconsin–Madison Lyric Bartholomay Iowa State University, [email protected] Hang-Yen Kou National Yang-Ming University Thomas A. Rocheleau University of Wisconsin - Madison Jeremy F. Fuchs UFonilvloerwsit ythi of sW aiscondn asiddn - itMionadisoaln works at: http://lib.dr.iastate.edu/ent_pubs Part of the Entomology Commons, Genetic Processes Commons, Genetics Commons, See next page for additional authors Genomics Commons, and the Laboratory and Basic Science Research Commons The ompc lete bibliographic information for this item can be found at http://lib.dr.iastate.edu/ ent_pubs/150. For information on how to cite this item, please visit http://lib.dr.iastate.edu/ howtocite.html. This Article is brought to you for free and open access by the Entomology at Iowa State University Digital Repository. It has been accepted for inclusion in Entomology Publications by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Construction and characterization of an expressed sequenced tag library for the mosquito vector Armigeres subalbatus Abstract Background The mosquito, Armigeres subalbatus, mounts a distinctively robust innate immune response when infected with the nematode Brugia malayi, a causative agent of lymphatic filariasis. In order to mine the transcriptome for new insight into the cascade of events that takes place in response to infection in this mosquito, 6 cDNA libraries were generated from tissues of adult female mosquitoes subjected to immune-response activation treatments that lead to well-characterized responses, and from aging, naïve mosquitoes. Expressed sequence tags (ESTs) from each library were produced, annotated, and subjected to comparative analyses. Results Six libraries were constructed and used to generate 44,940 expressed sequence tags, of which 38,079 passed quality filters to be included in the annotation project and subsequent analyses. All of these sequences were collapsed into clusters resulting in 8,020 unique sequence clusters or singletons. EST clusters were annotated and curated manually within ASAP (A Systematic Annotation Package for Community Analysis of Genomes) web portal according to BLAST results from comparisons to Genbank, and the Anopheles gambiae and Drosophila melanogaster genome projects. Conclusion The er sulting dataset is the first of its kind for this mosquito vector and provides a basis for future studies of mosquito vectors regarding the cascade of events that occurs in response to infection, and thereby providing insight into vector competence and innate immunity. Keywords Life Sciences, general, Microarrays, Proteomics, Animal Genetics and Genomics, Microbial Genetics and Genomics, Plant Genetics & Genomics Disciplines Entomology | Genetic Processes | Genetics | Genomics | Laboratory and Basic Science Research Comments This article is from BMC Genomics 8 (2007): 1-16, doi:10.1186/1471-2164-8-462. Rights This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. This article is available at Iowa State University Digital Repository: http://lib.dr.iastate.edu/ent_pubs/150 Authors George F. Mayhew, Lyric Bartholomay, Hang-Yen Kou, Thomas A. Rocheleau, Jeremy F. Fuchs, Matthew T. Aliota, I-Yu Tsao, Chiung-Yen Huang, Tze-Tze Liu, Kwang-Jen Hsiao, Shih-Feng Tsai, Ueng-Cheng Yang, Nicole T. Perna, Wen-Long Cho, Bruce M. Christensen, and Cheng-Chen Chen This article is available at Iowa State University Digital Repository: http://lib.dr.iastate.edu/ent_pubs/150 BMC Genomics BioMed Central Research article Open Access Construction and characterization of an expressed sequenced tag library for the mosquito vector Armigeres subalbatus George F Mayhew†1, Lyric C Bartholomay†2, Hang-Yen Kou3, Thomas A Rocheleau1, Jeremy F Fuchs1, Matthew T Aliota1, I-Yu Tsao3, Chiung-Yen Huang3, Tze-Tze Liu4, Kwang-Jen Hsiao5, Shih-Feng Tsai4,6, Ueng-Cheng Yang7, Nicole T Perna1, Wen-Long Cho3, Bruce M Christensen*1 and Cheng-Chen Chen*3 Address: 1Department of Pathobiological Sciences, University of Wisconsin-Madison, 1656 Linden Dr., Madison Wisconsin, 53706, USA, 2Department of Entomology, Iowa State University, Ames IA, 50010, USA, 3Department of Tropical Medicine, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan, 4VYM Genome Research Center, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan, 5Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei 112, Taiwan, 6Division of Molecular and Genomic Medicine, National Health Research Institutes, Miaoli County 350, Taiwan and 7Institute of Bioinformatics, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan Email: George F Mayhew - [email protected]; Lyric C Bartholomay - [email protected]; Hang-Yen Kou - [email protected]; Thomas A Rocheleau - [email protected]; Jeremy F Fuchs - [email protected]; Matthew T Aliota - [email protected]; I-Yu Tsao - [email protected]; Chiung-Yen Huang - [email protected]; Tze-Tze Liu - [email protected]; Kwang-Jen Hsiao - [email protected]; Shih- Feng Tsai - [email protected]; Ueng-Cheng Yang - [email protected]; Nicole T Perna - [email protected]; Wen- Long Cho - [email protected]; Bruce M Christensen* - [email protected]; Cheng-Chen Chen* - [email protected] * Corresponding authors †Equal contributors Published: 18 December 2007 Received: 19 September 2007 Accepted: 18 December 2007 BMC Genomics 2007, 8:462 doi:10.1186/1471-2164-8-462 This article is available from: http://www.biomedcentral.com/1471-2164/8/462 © 2007 Mayhew et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background: The mosquito, Armigeres subalbatus, mounts a distinctively robust innate immune response when infected with the nematode Brugia malayi, a causative agent of lymphatic filariasis. In order to mine the transcriptome for new insight into the cascade of events that takes place in response to infection in this mosquito, 6 cDNA libraries were generated from tissues of adult female mosquitoes subjected to immune-response activation treatments that lead to well- characterized responses, and from aging, naïve mosquitoes. Expressed sequence tags (ESTs) from each library were produced, annotated, and subjected to comparative analyses. Results: Six libraries were constructed and used to generate 44,940 expressed sequence tags, of which 38,079 passed quality filters to be included in the annotation project and subsequent analyses. All of these sequences were collapsed into clusters resulting in 8,020 unique sequence clusters or singletons. EST clusters were annotated and curated manually within ASAP (A Systematic Annotation Package for Community Analysis of Genomes) web portal according to BLAST results from comparisons to Genbank, and the Anopheles gambiae and Drosophila melanogaster genome projects. Conclusion: The resulting dataset is the first of its kind for this mosquito vector and provides a basis for future studies of mosquito vectors regarding the cascade of events that occurs in response to infection, and thereby providing insight into vector competence and innate immunity. Page 1 of 16 (page number not for citation purposes) BMC Genomics 2007, 8:462 http://www.biomedcentral.com/1471-2164/8/462 Background We previously reported a comprehensive analysis of ESTs The perpetuation of mosquito-borne diseases is depend- from complementary DNA (cDNA) libraries created from ent on the compatibility of the pathogen with its inverte- adult, female Ar. subalbatus hemocytes [1]. Experimental brate and vertebrate hosts, as dictated by each respective evidence has shown the importance of mosquito hemo- genome. The failure of traditional mosquito-borne dis- cytes (blood cells) as both initiators and mediators of ease control efforts to reduce the burden of these diseases mosquito immune responses [9-13]; therefore, material on public health has created an incentive to develop a was collected from the perfusate (which contains hemo- more comprehensive understanding of molecular interac- cytes) of Micrococcus luteus and Escherichia coli inoculated tions between host and pathogen, in order to develop mosquitoes at 1, 3, 6, 12, & 24 hours post bacterial inoc- novel means to control disease transmission. Innate ulation. These bacterial species have been extensively used immune responsiveness in the mosquito host is of partic- to examine immune peptide production in mosquitoes ular interest in such explorations because extensive [14,15], and each activates a different arm of the innate research efforts have shown that vector mosquito species immune response. The primary response of Ar. subalbatus produce robust humoral and cellular immune responses to E. coli is phagocytosis, whereas the primary response to against invading pathogens [1-4]. M. luteus is melanization, and it has been determined that this is independent of Gram type
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