THE CILIATE GENOMICS CONSORTIUM
CGC is a National Science Foundation funded consortium of undergraduate
student researchers and faculty members involved in the functional annotation
of genes in ciliated protozoans. So far the consortium has concentrated on
genes in Tetrahymena thermophila.
To study the function of Tetrahymena genes, research modules have been
developed for implementation as molecular biology class laboratory
exercises to involve a large number of undergraduate students in original
research.
The experimental results produced by students are published on a database
that is shared with the broader scientific community
(http://tet.jsd.claremont.edu)
Molecular Biology instructors can integrate these research modules into their
regular molecular laboratory class time to teach basic cloning and genetic
engineering techniques.
Consortium Research Modules
I. BIOINFORMATICS – A 1-2 week series of exercises to familiarize students with
basic gene structure and use of sequence databases to investigate putative gene
function. Oligonucleotides to use as primers in subsequent cloning molecules are
designed.
II. GENE TAG CONSTRUCTION (GTC) – A 11-13 week series of cloning exercises
that teach basic cloning and genetic engineering techniques in context of making
constructs that fuse genes of interest to common protein tags.
Tags
Green fluorescent protein (GFP) – for protein localization in live cells
Yellow fluorescent protein (YFP) – for protein localization in live cells
Cyan fluorescent protein (CFP) – for protein localization in live cells
2xHA – for protein localization in fixed cells by immunoflourescence,
and analysis of protein complexes by immunoprecipitation
FLAG-HIS6 – for purification of protein and/or analysis of protein
complexes
In this module, students will be able to transform their constructs into Tetrahymena
and visualize their tagged proteins by either direct fluorescence microscopy (GFP,
YFP, CFP tags) or by immunofluorescence/immunoblotting (2xHA & FLAG-HIS).
These activities can also be carried out by students in cell biology class laboratories.
III. KNOCKOUT CONSTRUCTION (KOC) – A 13-lab series of cloning exercises
applying basic cloning and genetic engineering techniques toward the generation
of gene knockout constructs.
Within this period of time students will be able to make a gene knockout
construct. Transformation of the construct and selection of the transformants is
performed outside of this module. Once transformants are obtained they can be
analyzed for phenotypes by future students in other classes such as cell biology.
IV. GENE EXPRESSION ANALYSIS (GE) – A 3-lab module in which students
analyze expression of a Tetrahymena gene throughout the life cycle, or in
response to different treatments.
THE TETRAHYMENA GENOMICS IN THE CLASSROOM
WORKSHOP
This workshop brings together undergraduate teaching faculty from a variety
of institutions to learn how to integrate and implement the Tetrahymena
genomics research modules in their classrooms. The hope is that this
workshop will provide the practical foundation, collegial support, and
materials necessary to increase the number of undergraduates engaging
original research on the function of Tetrahymena genes at the participating
institutions. Students and faculty involved in this research effort may publish
their experimental results on a common database, which is accessible to the
broader scientific community, and which facilitates scientific communication
between members of the consortium.