Archived Abstracts

Poster or Presentation Title

Constructing Plasmids for Inducible Expression aCd Purification of Colorful Recombinant Chromoproteins in Escherichia Coli

Student Author Information

Janaya Mott, University of LynchburgFollow

Location

Memorial Ballroom, Hall Campus Center

Access Type

Open Access

Presentation Type

Poster Presentation

Start Date

8-4-2020 12:00 PM

End Date

8-4-2020 1:15 PM

Department

Biology

Abstract

Many living organisms can naturally produce vivid colors, sometimes through the expression of chromoproteins. Chromoproteins are complex proteins that partially consist of pigmented cofactors. These proteins can be used to alter the genome of bacteria causing them to grow in a specific color depending on the chromoprotein used. Our objective is to create four different inducible color Escherichia coli strains using the chromoproteins scOrange, gfasPurple, aeBlue, and amajLime. Each chromoprotein will be inserted into the multiple cloning site of a pET-15b plasmid vector. The pET-15b vector contains an ampicillin resistance gene, histidine-tag with six histidines, and a lac repressor controlled by Isopropyl-β-D-thiogalactoside (IPTG) signaling. Results are supposed to show a strain of E. coli that can be IPTG-induced to grow in the color determined by the chromoprotein that it was transformed with. The histidine-tagged chromoprotein can then be purified by affinity chromatography. The results that were obtained from this experiment showed that the gfaspurple was properly induced into the genome but stressed under IPTG induction, no color was expressed. The ultimate goal of this work is to create colorful molecular tools that are affordable and suitable for high school or undergraduate students that are interested in practicing microbiology or learning the central dogma. The characteristics of these recombinant vectors will allow us to clearly visualize bacterial transformation and use an engaging and colorful method to visualize the process of gene expression.

Faculty Mentor(s)

Dr. Jamie Brooks

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Apr 8th, 12:00 PM Apr 8th, 1:15 PM

Constructing Plasmids for Inducible Expression aCd Purification of Colorful Recombinant Chromoproteins in Escherichia Coli

Memorial Ballroom, Hall Campus Center

Many living organisms can naturally produce vivid colors, sometimes through the expression of chromoproteins. Chromoproteins are complex proteins that partially consist of pigmented cofactors. These proteins can be used to alter the genome of bacteria causing them to grow in a specific color depending on the chromoprotein used. Our objective is to create four different inducible color Escherichia coli strains using the chromoproteins scOrange, gfasPurple, aeBlue, and amajLime. Each chromoprotein will be inserted into the multiple cloning site of a pET-15b plasmid vector. The pET-15b vector contains an ampicillin resistance gene, histidine-tag with six histidines, and a lac repressor controlled by Isopropyl-β-D-thiogalactoside (IPTG) signaling. Results are supposed to show a strain of E. coli that can be IPTG-induced to grow in the color determined by the chromoprotein that it was transformed with. The histidine-tagged chromoprotein can then be purified by affinity chromatography. The results that were obtained from this experiment showed that the gfaspurple was properly induced into the genome but stressed under IPTG induction, no color was expressed. The ultimate goal of this work is to create colorful molecular tools that are affordable and suitable for high school or undergraduate students that are interested in practicing microbiology or learning the central dogma. The characteristics of these recombinant vectors will allow us to clearly visualize bacterial transformation and use an engaging and colorful method to visualize the process of gene expression.