Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1998 Mar 31;95(7):3743-7.
doi: 10.1073/pnas.95.7.3743.

Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly

N Jasinskiene  1 , C J Coates, M Q Benedict, A J Cornel, C S Rafferty, A A James, F H Collins
Affiliations

Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly

N Jasinskiene et al. Proc Natl Acad Sci U S A. .

Abstract

The mosquito Aedes aegypti is the world's most important vector of yellow fever and dengue viruses. Work is currently in progress to control the transmission of these viruses by genetically altering the capacity of wild Ae. aegypti populations to support virus replication. The germ-line transformation system reported here constitutes a major advance toward the implementation of this control strategy. A modified Hermes transposon carrying a 4.7-kb fragment of genomic DNA that includes a wild-type allele of the Drosophila melanogaster cinnabar (cn) gene was used to transform a white-eyed recipient strain of Ae. aegypti. Microinjection of preblastoderm mosquito embryos with this construct resulted in 50% of the emergent G0 adults showing some color in their eyes. Three transformed families were recovered, each resulting from an independent insertion event of the cn+-carrying transposon. The cn+ gene functioned as a semidominant transgene and segregated in Mendelian ratios. Hermes shows great promise as a vector for efficient, heritable, and stable transformation of this important mosquito vector species.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Eye color phenotypes resulting from the G1 intercross of family 64. Animals were photographed with incident light under a dissecting microscope. (Upper Left) khw recipient strain white eye phenotype. (Upper Right) Wild-type eye color. Transformed members of family 64 carrying two copies (Lower Right) and one copy (Lower Left) of an insertion.
Figure 2
Figure 2
(A) A representational map of the partial structure of the Hermes construct, pH[cn]. The Hermes left (LH) and right (RH) inverted terminal repeats (arrows) flank the 4.7-kb genomic fragment of D. melanogaster DNA carrying a wild-type copy of the cinnabar gene (cn+) (boxed region). This potentially mobile portion of the pH[cn] construct is flanked by M. domestica genomic DNA (MD, dotted lines). The positions of two restriction endonuclease-cleavage sites (SacII and XbaI) are indicated. The relative extents and sizes of the DNA fragments used as probes in the Southern analyses are shown below the map (numbers and solid lines). (B) Southern analysis of genomic DNA isolated from G1 progeny of transformed families 35 and 64, and the control recipient strain, khw, digested with XbaI, which cuts once in the integrated fragment, hybridized with the 0.76-kb LH-labeled fragment. Unique hybridization signals associated with insertions in families 35 and 64 are seen at 12 and 11 kb, respectively. Lower molecular weight signals corresponding to cross-hybridizing fragments are seen in the transformed families and recipient strain. (C) Southern blot analysis of genomic DNA isolated from G2 mosquitoes of families 35, 64, and the pool (P), and the recipient strain digested with either XbaI or XbaI/SacII, and hybridized with the 4.7-kb cn+ labeled fragment. High molecular weight signals corresponding to fragment lengths of 12, 11, and 10 kb are seen in the XbaI digests of DNA from families 35, 64, and the pool, respectively. The expected 4.7-kb fragment size is evident in the XbaI/SacII digest of DNA from family 64 and the pool. However, a truncated signal at 4.5 kb is seen in family 35 DNA, indicating loss of some DNA. (D) Southern blot analysis of genomic DNA isolated from G2 mosquitoes of families 35, 64, and the recipient strain, khw digested with XbaI, and hybridized with the MD-labeled fragments. The positive control is XbaI-digested pH[cn].
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Collins F H, James A A. Sci Med. 1996;3:52–61.
    1. Meredith S E O, James A A. Ann Parasitol Humaine Comp. 1990;65:S1:113–118. - PubMed
    1. Miller L H, Sakai R K, Romans P, Gwadz R W, Kantoff P, Coon H G. Science. 1987;237:779–781. - PubMed
    1. McGrane V, Carlson J O, Miller B R, Beaty B J. Am J Trop Med Hyg. 1988;39:502–510. - PubMed
    1. Morris A C, Eggleston P, Crampton J M. Med Vet Entomol. 1989;3:1–7. - PubMed

Publication types

Substances

LinkOut - more resources