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LOGO: Journal of Cotton Science


Detection of Enhanced Green Fluorescent Protein DNA in Pink Bollworm through Polymerase Chain Reaction

Authors: John J. Peloquin and T. A. Miller
Pages: 28-33
Molecular Biology and Physiology

Indelible marking of pink bollworm (Pectinophora gossypiella) would assist in implementation of sterile insect technique-based pest control of this insect. Additionally, genetic manipulation through DNA-mediated genetic transformation also requires a reliable genetic marker. To this end, we injected DNA plasmids encoding the enhanced green fluorescent protein gene into pink bollworm pre-blastoderm embryos and analyzed in-vivo the expression of DNA that encodes the enhanced green fluorescent protein. As a confirmatory technique for detection of the protein-encoding DNA, in addition to visual detection of the protein, we used the polymerase chain reaction (PCR) to amplify the expected 579 bp enhanced green fluorescent protein DNA fragment from DNA extracted from green fluorescent protein-positive pink bollworm. We did not amplify such a 579 bp DNA fragment from negative control DNA templates or from DNA isolated from pink bollworm that did not exhibit green fluorescence from the enhanced protein.