The goal of this project is to develop mariner transposable elements as genetic tools for arthropods, primarily insects, of medical importance. The impact of arthropod vectors of disease around the world is enormous, and novel approaches to their control are needed. These mariner transposons are potentially useful for gene tagging for cloning, for enhancer trapping, and most importantly as transformation vectors, both for basic research and practical approaches to mitigating transmission of arthropod-borne diseases. There is an extraordinary diversity of mariner family transposons in the genomes of diverse animals. They appear to function independently of host specific factors, and therefore any particular mariner transposon is a candidate for development. A particular element called Himar1 has been extensively studied by our laboratory, and shown to be active in vitro and in vivo in Drosophila melanogaster. This activity is, however, limited by a concentration- dependent inhibition of the mariner transposase, which appears to be a general feature of mariner transposons and may be shared with some bacterial transposons such as Tn5. We propose to overcome this inhibition by isolating hyperactive uninhibited versions of Himar1 using a mutagenic screen in bacteria, such as E. coli. These will be tested in vitro and then in vivo in D. melanogaster, where they will be developed into transformation vectors, followed by distribution to colleagues for testing in diverse insects of medical importance. Additional hyperactive versions of Himar1 might be obtained by selecting for changes in their termini and flanking DNA. The fate of Himar1 transposase within insect cells and embryos will be examined to determine whether it can be stabilized. The interactions of Himar1 transposase with other mariners of diverse subfamilies will be examined to evaluate whether Himar1 will interact with endogenous mariners in host genomes. This project will hopefully lead to the availability of an efficient general transformation system for diverse arthropods of medical, economical, and basic research importance, and may also lead to development of genetic tools in other organisms.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI033586-06
Application #
6169946
Study Section
Special Emphasis Panel (ZRG5-TMP (01))
Program Officer
Aultman, Kathryn S
Project Start
1994-01-01
Project End
2002-07-31
Budget Start
2000-08-01
Budget End
2002-07-31
Support Year
6
Fiscal Year
2000
Total Cost
$161,987
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Zoology
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Barry, Elizabeth G; Witherspoon, David J; Lampe, David J (2004) A bacterial genetic screen identifies functional coding sequences of the insect mariner transposable element Famar1 amplified from the genome of the earwig, Forficula auricularia. Genetics 166:823-33
Lampe, David J; Witherspoon, David J; Soto-Adames, Felipe N et al. (2003) Recent horizontal transfer of mellifera subfamily mariner transposons into insect lineages representing four different orders shows that selection acts only during horizontal transfer. Mol Biol Evol 20:554-62
Lampe, D J; Walden, K K; Robertson, H M (2001) Loss of transposase-DNA interaction may underlie the divergence of mariner family transposable elements and the ability of more than one mariner to occupy the same genome. Mol Biol Evol 18:954-61
Zhang, J K; Pritchett, M A; Lampe, D J et al. (2000) In vivo transposon mutagenesis of the methanogenic archaeon Methanosarcina acetivorans C2A using a modified version of the insect mariner-family transposable element Himar1. Proc Natl Acad Sci U S A 97:9665-70
Lampe, D J; Akerley, B J; Rubin, E J et al. (1999) Hyperactive transposase mutants of the Himar1 mariner transposon. Proc Natl Acad Sci U S A 96:11428-33
Rubin, E J; Akerley, B J; Novik, V N et al. (1999) In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria. Proc Natl Acad Sci U S A 96:1645-50
Lampe, D J; Grant, T E; Robertson, H M (1998) Factors affecting transposition of the Himar1 mariner transposon in vitro. Genetics 149:179-87
Robertson, H M; Lampe, D J (1995) Recent horizontal transfer of a mariner transposable element among and between Diptera and Neuroptera. Mol Biol Evol 12:850-62
Robertson, H M; Lampe, D J (1995) Distribution of transposable elements in arthropods. Annu Rev Entomol 40:333-57