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Submitted to: Insect Molecular Biology
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 10/29/1996 Publication Date: N/A Citation: N/A Interpretive Summary: The ability to achieve gene transfer in economically important insects depends upon an understanding of transposon gene vector and marker system functions in host insects. In this report scientists at the Insect Attractant Laboratory in Gainesville, Florida describe results from experiments designed to test the ability of the hobo gene-transfer vector from Drosophila melanogaster to mediate gene-transfer in a distantly related drosophilid species, D. virilis, as a prelude to testing it in more distantly related nondrosophilid insects. As part of this experiment the ability of the D. melanogaster white+ eye-color gene to act as a "mutant rescue" marker in a white eye-color mutant strain of D. virilis was tested. Using a vector containing the white+ gene within a hobo element, a single transformant was identified (at an approximately 0.5% frequency) having a strong red eye color phenotype, indicating that the hobo vector is functional in D. virilis but at a reduced level compared to D. melanogaster. Importantly, the D. melanogaster white+ gene may be a useful marker system for various fruit fly and moth species having white mutant strains. Thus, this work provides a background for genetic manipulation of pest insects. Technical Abstract: A Drosophila virilis white mutant host strain was transformed with a hobo vector containing the D. melanogaster mini-white+ cassette (H[w+, hawN]). Transformation was catalyzed by an unmodified hobo transposase helper (pHFL1). Of 600 injected embryos, 265 G0 adults were intermated in 73 lines. An intermediate to strong red-eye color phenotype was observed in five G1 individuals from the same G0 intermated line. Southern hybridization and chromosomal in situ hybridization indicated a single event had occurred. Assuming a 70% fertility rate, an approximate transformation frequency of approximately 0.5% is deduced. Crosses indicated X-chromosome linkage which was verified by chromosomal hybridization which further localized integration to position 19A/B. Several wing and eye morphology mutants were observed in both G0 and G1 individuals (in 13 separate lines) suggesting that hobo may have mobilized a related element in D. virilis. The data indicate function of both the hobo transposon vector and white+ marker from D. melanogaster in a distantly related drosophilid suggesting possible function in nondrosophilid species as well. |
