Western Kentucky University

PiRaTe-PYG: A New Genetic Construct for Studying Gene Expression in Butterflies

Institution

Western Kentucky University

Abstract

We are constructing a DNA construct to inject into butterflies. This construct will integrate into the DNA of the butterfly, creating mutations, and allow us to study how butterflies make color patterns on their wings. This construct can also be used in other insects to study a variety of genetic processes. The construct contains several elements. First, it contains the inverse repeats of the transposable element piggyBac, a “jumpinggene” that moves from place to place in the genome of an organism. Second, it contains two traceable markers—green fluorescent protein and red fluorescent protein, which allow the researcher to trace the construct once it is introduced into the animal. Third, it contains a gene that encodes the transponase enzyme that allows the construct to integrate into the DNA of the animal. The construct is designed to break apart after it is introduced into the animal to produce two useful halves for further research. Finally, it contains a gene that allows the construct to “borrow” regulatory elements from a nearby butterfly gene, so that green fluorescence is expressed under the same conditions as the nearby butterfly gene. This will allow us to visualize using fluorescent microscopy the changing patterns of gene expression associated with color pattern development as the wing develops in real time. This will allow us to study butterfly wings as a general model for cell growth and differentiation.

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PiRaTe-PYG: A New Genetic Construct for Studying Gene Expression in Butterflies

We are constructing a DNA construct to inject into butterflies. This construct will integrate into the DNA of the butterfly, creating mutations, and allow us to study how butterflies make color patterns on their wings. This construct can also be used in other insects to study a variety of genetic processes. The construct contains several elements. First, it contains the inverse repeats of the transposable element piggyBac, a “jumpinggene” that moves from place to place in the genome of an organism. Second, it contains two traceable markers—green fluorescent protein and red fluorescent protein, which allow the researcher to trace the construct once it is introduced into the animal. Third, it contains a gene that encodes the transponase enzyme that allows the construct to integrate into the DNA of the animal. The construct is designed to break apart after it is introduced into the animal to produce two useful halves for further research. Finally, it contains a gene that allows the construct to “borrow” regulatory elements from a nearby butterfly gene, so that green fluorescence is expressed under the same conditions as the nearby butterfly gene. This will allow us to visualize using fluorescent microscopy the changing patterns of gene expression associated with color pattern development as the wing develops in real time. This will allow us to study butterfly wings as a general model for cell growth and differentiation.