Title

Expression and Purification of Neocortical Arealization Factors

Presenter Information

Alonna Miller
Nick Meyers

Document Type

Oral Presentation

Location

SURC Ballroom C/D

Start Date

16-5-2013

End Date

16-5-2013

Abstract

The neocortex is a mammalian-specific division of the cerebral cortex involved in higher cognitive functions including conscious thoughts and actions. The neocortex is divided into discrete functional areas that process specific types of visual, auditory, motor, and somatosensory (touch) information. The embryonic process by which these different neocortical areas take on their functional properties is called arealization. This process is mediated by a specific class of proteins called transcription factors; proteins that determine when genes are turned on or off. In order to reveal how one of these transcription factors, Emx2, mediates neocortical arealization, we performed a genetic screen to identify proteins that are potentially interacting with Emx2. The next phase of this research will be to use a biochemical experiment, co-immunoprecipitation, to confirm these protein-protein interactions in a much more stringent manner. The first step in conducting a co-immunoprecipitation is the generation and purification of the interacting proteins. The DNA sequences coding for each of the interacting proteins have been cloned into bacterial expression plasmids that will generate our proteins with additional protein tags that will be used for purification. We have successfully used these plasmids to program E. coli to produce high levels of the desired proteins. Our work is currently focused on utilizing the “tags” on our recombinant proteins for affinity purification, which is the last step required before they can be used for in vitro co-immunoprecipitation assays.

Poster Number

41

Faculty Mentor(s)

Todd Kroll

Additional Mentoring Department

Chemistry

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May 16th, 8:20 AM May 16th, 10:50 AM

Expression and Purification of Neocortical Arealization Factors

SURC Ballroom C/D

The neocortex is a mammalian-specific division of the cerebral cortex involved in higher cognitive functions including conscious thoughts and actions. The neocortex is divided into discrete functional areas that process specific types of visual, auditory, motor, and somatosensory (touch) information. The embryonic process by which these different neocortical areas take on their functional properties is called arealization. This process is mediated by a specific class of proteins called transcription factors; proteins that determine when genes are turned on or off. In order to reveal how one of these transcription factors, Emx2, mediates neocortical arealization, we performed a genetic screen to identify proteins that are potentially interacting with Emx2. The next phase of this research will be to use a biochemical experiment, co-immunoprecipitation, to confirm these protein-protein interactions in a much more stringent manner. The first step in conducting a co-immunoprecipitation is the generation and purification of the interacting proteins. The DNA sequences coding for each of the interacting proteins have been cloned into bacterial expression plasmids that will generate our proteins with additional protein tags that will be used for purification. We have successfully used these plasmids to program E. coli to produce high levels of the desired proteins. Our work is currently focused on utilizing the “tags” on our recombinant proteins for affinity purification, which is the last step required before they can be used for in vitro co-immunoprecipitation assays.