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A Microbiological and Bioinformatics Safari: Searching for Cellulolytic Prokaryotes from the Environment and Characterization of their Cellulase Genes.

Cellulases are enzymes that digest the common polysaccharide cellulose using one of several enzymatic strategies (EC 3.2.1.4). A variety of organisms, ranging from marine to terrestrial, aerobic to anaerobic, possess enzymes with this activity. Cellulases may prove to be of significant economic importance to the process of converting cellulose to ethanol, in other words, the burgeoning biofuel industry. We propose a multi-institutional effort to identify cellulolytic organisms by enrichment, create transposon mutants lacking cellulase activity (via screening of mutants), obtain DNA sequence from these mutants, and to compare the recovered cellulase-specific gene sequences to known cellulase genes. The 16S rRNA sequence from each cellulolytic isolate will also be obtained, and the phylogenetics of the organism and the cellulases will be examined and compared. Cellulase specific oligonucleotide primers can be designed to locate cellulase sequences from microbially diverse environments without cultivation and mutagenesis. Students can also search publically-available metagenomic data sets for cellulase sequences to explore cellulase diversity from geographically distinct environments. Such sequences will provide useful information regarding the diversity and phylogeny of cellulase-like genes from a variety of environments. Finally research on the regulation of cellulase can be investigated on the isolated strains. Different institutions will carry out these procedures from a variety of environments, such as marine sediments, estuaries, and agricultural soils. The strains, mutants, and sequences recovered will be a resource available to participating investigators and their students. A website will be maintained for easy access to data and current investigations. Students will also present their data at annual group meetings among the participating institutions.

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Resource Type: Assignment/Activity, Teaching strategies & guidelines
Audience Level: Undergraduate upper division 15-16, Undergraduate lower division 13-14

Author and Copyright


Authors and Editors: Anne Bernhard of Connecticut College, Mark Martin of University of Puget Sound, Shannon Hinsa-Leasure of Michigan State University
Publisher: BioQUEST Curriculum Consortium
Format: text/html
Copyright and other restrictions: No
Cost: No

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