Formation and loss of large, unstable tandem arrays of the piggyBac transposable element in the yellow fever mosquito, Aedes aegypti.
The Class II transposable element, piggyBac, was used to transform the yellow fever mosquito, Aedes aegypti. In two transformed lines only 15-30% of progeny inherited the transgene, with these individuals displaying mosaic expression of the EGFP marker gene. Southern analyses, gene amplification of genomic DNA, and plasmid rescue experiments provided evidence that these lines contained a high copy number of piggyBac transformation constructs and that much of this DNA consisted of both donor and helper plasmids. A detailed analysis of one line showed that the majority of piggyBac sequences were unit-length donor or helper plasmids arranged in a large tandem array that could be lost en masse in a single generation. Despite the presence of a transposase source and many intact donor elements, no conservative (cut and paste) transposition of piggyBac was observed in these lines. These results reveal one possible outcome of uncontrolled and/or unexpected recombination in this mosquito, and support the conclusion that further investigation is necessary before transposable elements such as piggyBac can be used as genetic drive mechanisms to move pathogen-resistance genes into mosquito populations.
Adelman
Zach N
ZN
Department of Molecular Biology and Biochemistry, University of California, Irvine, CA 92697-3900, USA.
Jasinskiene
Nijole
N
Vally
K J M
KJ
Peek
Corrie
C
Travanty
Emily A
EA
Olson
Ken E
KE
Brown
Susan E
SE
Stephens
Janice L
JL
Knudson
Dennis L
DL
Coates
Craig J
CJ
James
Anthony A
AA
D000330:Aedes / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D016367:Baculoviridae / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics; D017344:Genes, Insect; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D016254:Mutagenesis, Insertional; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins