A new dominant selectable marker for genetic transformation; Hsp70-opd

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• P element plasmids are crucial for Drosophila melanogaster transformation.
• The organophosphate-degrading gene (opd) offers a potential selectable marker.
• The mini-white gene is a commonly used comarker in Drosophila transformation.

Purpose of the Study:

• To evaluate new P element plasmids containing the opd gene as transformation vectors in Drosophila melanogaster.
• To assess the efficacy of the Hsp70-opd gene as a dominant selectable marker.
• To compare the Hsp70-opd marker with the mini-white gene for identifying transformed individuals.

Main Methods:

• Construction and testing of P element plasmids with the opd gene.
• Modification of a vector by adding the D. melanogaster mini-white gene.
• Identification of transformed individuals using paraoxon selection for opd.
• Evaluation of Hsp70-opd and mini-white for distinguishing transformed individuals.

Main Results:

• Transformed individuals identified using paraoxon selection for opd showed results similar to those obtained with mini-white.
• No false positives were recovered.
• One strain with the mini-white gene had inadequate resistance due to a defective Hsp70-opd gene.
• Hsp70-opd proved similar to mini-white for distinguishing transformed individuals without time-consuming examination.

Conclusions:

• The Hsp70-opd gene is an effective dominant selectable marker in Drosophila melanogaster.
• Hsp70-opd offers an efficient alternative to mini-white, reducing screening time.
• The Hsp70-opd marker has potential applications in various animal models due to its mode of action against organophosphorus nerve agents.