Abstract
Characterizing symbiotic communities, like that of the termite hindgut, is essential for understanding their functionality and capabilities. However, the same complexity that allows termites to digest wood so efficiently also makes them difficult to study. With the expansion in technology and sequencing strategies the feasibility of sequencing entire consortiums or microecosystems is now possible. Here we present an adapted library preparation strategy which allows for the detection and measurement of expressed genes from all three domains of life in a single sample simultaneously. This technique effectively captures the transcriptome contributions by the various members of the consortium regardless of their taxonomic identity, which can then be annotated using custom-built databases and reciprocal BLASTing. Joining the universality of this library prep strategy with the power of bioinformatics allows for the identification of cellulases and other genes encoding carbohydrate active enzymes from complex communities using metatranscriptomics.
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Acknowledgments
The authors acknowledge Dr. Zul Gulzar and technicians at NuGen Technologies for helpful consultation and design of the custom InDA-C oligos. We also thank Steve Kelley, Michael Gribskov, and Pete Pascuzzi for invaluable discussion, training, and guidance in the development of the annotation strategies detailed herein. Funding support for this work was provided by the O. Wayne Rollins-Orkin Endowment in the Department of Entomology at Purdue University, the Monsanto Research Grant from the Entomological Society of America, the Indiana Academy of Sciences Senior Research Grant (Grant #: 2014-13), and the Postdoctoral Excellence in Research and Teaching Fellowship administered by the Center for Insect Science at the University of Arizona funded through the NIH-IRACDA program (Grant #: K12GM000708-17).
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Peterson, B.F., Scharf, M.E. (2018). Metatranscriptomic Techniques for Identifying Cellulases in Termites and their Symbionts. In: Lübeck, M. (eds) Cellulases. Methods in Molecular Biology, vol 1796. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7877-9_7
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DOI: https://doi.org/10.1007/978-1-4939-7877-9_7
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