David Lampe, Ph.D.Associate Proessor
Bayer School of Natural and Environmental Sciences
209 Mellon Hall
Education:Ph.D., Entomology, University of Illinois at Urbana-Champaign, 1992
M.S., Entomology, Purdue University, 1987
B.S., Biology, St. Louis University, 1985
BIOL112 Evolution, Ecology, and Diversity
BIOL417/517 Invertebrate Biology and Biotechnology
I. Symbiotic control of insect transmitted diseases of plants and animals.
Insects are vectors for many of the most deadly plant and animal diseases and are a crucial link in these disease cycles. In the past, cultural methods or insecticide treatments have been used to control insect vectors. These methods are still widely practiced, but in many cases insects have evolved resistance to insecticides. In some areas of the world, notably Africa, public health measures used in developed countries to reduce vector populations are difficult to apply. Clearly, new methods are necessary to aid in the control of insect-vectored diseases.
Symbiotic control is a method that takes advantage of basic microbial ecology. The phenotype of all plants and animals is the product of genetic and environmental effects. One large environmental effect is provided by the microorganisms that form various kinds of sybioses with plants and animals. In symbiotic control, we genetically engineer symbiotic microbes to deliver -effector- proteins that can interfere with disease causing organisms. In this way, we can alter the disease-causing phenotype of insect vectors indirectly by modifying the bacteria that they normally carry.
1. Blocking mosquito transmission of malaria to humans. In a collaboration with Marcelo Jacobs-Lorena at Johns Hopkins University, we are genetically modifying two species of bacteria to deliver antimalarial effector proteins. Malaria is the most widespread and dangerous insect-transmitted human disease in the world. It infects more than 500 million people (ca. 1 in 12 humans) and causes between 1 and 2 million deaths each year. The incidence of malaria is increasing and new measures to combat it are desperately needed.
Both Pantoea agglomerans and Asaia borogensis are bacterial species that are found in the midguts of Anopheles gambiae, the most important malaria vector mosquito in Africa. We are developing secretion systems for use in both of these species in order to secrete anti-malarial effectors into the midgut of An. gambiae. These effector molecules include single chain antibodies that bind directly to the Plasmodium parasites that cause malaria and to receptors on the mosquito midgut epithelium that the parasite uses to invade mosquito tissues.
2. Blocking leafhopper transmission of Pierce's Disease to grapevines. Insects also transmit diseases to plants. In California, the glassy-winged sharpshooter (a leafhopper) is the most important vector of the bacterium, Xylella fastidiosa. When this bacterium infects the xylem of grapevines (especially Vitis vinifera, the grape species used to make win) it blocks water flow in the plants leading to a condition called Pierce's Disease, which is nearly always fatal to the plant. Very costly measures are used to keep the sharpshooters under control and thus keep the incidence of Pierce's Disease to manageable levels.
We are developing a symbiotic control strategy in order to prevent the spread of Pierce's Disease and also to help cure infected grapevines. We have isolated single chain antibodies that bind specifically to the surface of Xylella and these were recently shown to drastically reduce the ability of sharpshooters to transmit Xyella to non-infected plants. We are collaborating with Angray Kang of Westminster University in the UK to engineer these antibodies into forms that will be able to be secreted from bacteria that live in both sharpshooters and grapevine xylem. We are also developing single chain antibodies directed against polygalacturonase, an enzyme Xylella secretes in order to colonize the plant. Finally, we are engineering polygalacturonase inhibitor protein from Bartlett pears for the ability to be secreted by symbiotic bacteria.
II. Molecular biology and evolution of mariner transposable elements
The other main focus of my lab is the biology of a family of transposons known as mariners or mariner-like elements. All organisms contain transposable elements of various kinds. These are genes that occupy no necessarily-fixed location in the genome as do other genes. Instead, transposable elements move around the genome and as a consequence are able to make copies of themselves. In doing so, they can do real harm to the host organism by causing chromosomal breakage and gene inactivation through insertion into those genes. Hence, there is a balance that the transposon needs to make between high activity (which would kill the host) and no activity (which would mean elimination of the transposon from the genome). In a very real sense, these genes are the pinnacle of selfish DNA.
The primary goal of our transposon research is to understand how a particular mariner, the Himar1 element from the horn fly, Haematobia irritans, functions on the biochemical and genetic levels and how these elements evolve. To this end, we have reconstituted the transposition of this element in vitro and are actively investigating how the transposase protein encoded by Himar1 is able to perform all of the functions necessary to move its cognate gene from one DNA molecule to another. A fundamental understanding of how this occurs will allow us to better understand the evolution of these genes and to create efficient genetic tools from them for use in microorganisms, insects, and vertebrates.
Symbiotic control and insect transgenesis.
- Azizi A, Arora A, Markiv A, Lampe DJ, Miller TA, Kang AS. 2012. Ribosome display of combinatorial antibody libraries derived from mice immunized with heat-killed Xylella fastidiosa and the selection of MopB-specific single-chain antibodies. Appl Environ Microbiol. 78(8):2638-47.
- Bisi DC, Lampe DJ. 2011. Secretion of anti-Plasmodium effector proteins from a natural Pantoea agglomerans isolate by using PelB and HlyA secretion signals. Appl Environ Microbiol. 77(13):4669-75.
- Riehle MA, Moreira CK, Lampe D, Lauzon C, and Jacobs-Lorena, M. 2007. Using bacteria to express and display anti-Plasmodium molecules in the mosquito midgut. International Journal of Parasitology 37: 596-603.
- Miller, T. A., C. R. Lauzon, D. Lampe, R. Durvasula and C. Matthews. 2006. 'Paratransgenesis applied to control insect-transmitted plant pathogens: The Pierce's disease case-. In: Insect Symbiosis 2 K. Bourtzis and T. A. Miller, eds. Taylor and Francis, London/CRC Press Boca Raton, FL
- Miller, Thomas A., David J. Lampe and Carol R. Lauzon. 2006 -Transgenic and paratransgenic insects in crop protection.' In: Insecticide Design Using Advanced Technologies. Isaac Ishaaya, Ralf Nauen and Rami Horowitz, eds. Springer-Verlag, Heidelberg, Germany.
- Bextine B, Lampe D, Lauzon C, Jackson B, Miller TA. 2005. Establishment of a Genetically Marked Insect-Derived Symbiont in Multiple Host Plants. Current Microbiology 50: 1-7.
- Bextine, B.R., Lauzon, C.R., Potter, S.E., Lampe, D. and Miller, T.A. 2004. Delivery of a genetically marked Alcaligenes sp. to the glassy-winged sharpshooter for use in a paratransgenic control strategy. Current. Microbiolology 48: 327-331.
- Lampe, D.J., Walden, K.K.O., Sherwood, J.M., and Robertson, H.M. 2000. "Genetic engineering with mariner transposons". In. Insect Transgenesis: Methods and Applications, A. Handler and D. O'Brochta, eds. CRC Press.
- Rubin, E.J., Akerley, B.J., Lampe, D.J., Husson, R.N., and Mekalanos, J.J. 1999. In vivo transposition of mariner-based elements in enteric bacteria and mycobacteria. Proceedings of the National Academy of Sciences 96: 1645-1650.
Mariner transposable elements
- Lampe, D.J. 2010. Bacterial genetic methods to explore the biology of mariner transposons. Genetica, 138(5):499-508.
- Keravala, A. , Liu, D., Lechman, E.R., Wolfe, D., Nash, J., Lampe, D.J., and Robbins, P.D. 2006. Hyperactive Himar1 Transposase Mediates Transposition In Cell Culture And Enhances Gene Expression In Vivo. Human Gene Therapy 17: 1006-1018.
- Butler, M.G., Chakraborty, S., and Lampe, D.J. 2006. The N-terminus of Himar1 mariner transposase mediates multiple activities during transposition. Genetica, 127(1-3):351-66.
- Barry, E. G., Witherspoon, D.J., and Lampe, D.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: 823-833.
- Lipkow K., Buisine N., Lampe D.J., Chalmers R. 2004. Early intermediates of mariner transposition: catalysis without synapsis of the transposon ends
- uggests a novel architecture of the synaptic complex. Molecular and Celular Biology 24: 8301-8311.
- Lampe D.J., Witherspoon D.J., Soto-Adames F.N., Robertson H.M. 2003 Recent horizontal transfer of mellifera subfamily mariner transposons into insect lineages representing four different orders shows that selection acts only during horizontal transfer. Molecular Biology and Evolution 20:554-62
- Akerley, B.J. and Lampe, D.J. 2002. The GAMBIT system for analysis of virulence and essential genes. Methods in Enzymology 358:100-8.
- Lampe, D. J., K. K. O. Walden and H. M. Robertson 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. Molecular Biology and Evolution 18: 954-961.
- Hamer, L., Woessner, J., Montenegro-Chamorro, M.V., Adachi, K., Tarpey, R.W., Lampe, D.J., Slater, T., Ramamurthy, L., and Hamer, J.E. 2001. Gene discovery and gene function assignment in filamentous fungi. Proceedings of the National Academy of Sciences 98:5110-5115.
- Pelicic, V., Morelle, S., Lampe, D., Nassif, X. 2000. Mutagenesis of Neisseria meningitidis by in vitro transposition of Himar1 mariner. Journal of Bacteriology 182:5391-8.
- Zhang, J.K., Pritchett, M.A., Lampe, D.J., Robertson, H.M., Metcalf, W.W. 2000. In vivo transposon mutagenesis of the methanogenic archaeon Methanosarcina acetivorans C2A using a modified version of the insect mariner-family transposable element Himar1. Proceedings of the National Academy of Sciences 97:9665-70.
- Lampe, D. J., Akerley, B.J., Rubin, E. J., Mekalanos, J.J., and Robertson, H.M. 1999 Hyperactive mutants of Himar1 mariner transposase. Proceedings of the National Academy of Sciences 96:11428-11433.
- Zhang, L., Sankar, U., Lampe, D.J., Robertson, H.M., and Graham, F.L.. 1998. The Himar1 mariner transposase encoded by a recombinant Ad vector is functional in mammalian cells. Nucleic Acids Research, 26: 3687-3693.
- Akerley, B.J., Rubin, E.J., Camilli, A., Lampe, D.J., Robertson, H.M., and Mekalanos, J.J. 1998. Systematic identification of essential genes by in vitro Himar1 mariner mutagenesis. Proceedings of the National Academy of Sciences 95: 8927-8932.
- Lampe, D.J., Grant, T.E., and Robertson, H.M. 1998. Factors affecting transposition of the Himar1 mariner transposon in vitro, Genetics 149: 179-187.
- Lampe, D.J., Churchill, M.E.A., and Robertson, H.M. 1996. A purified mariner transposase is sufficient to mediate transposition in vitro. European Molecular Biology Journal 15: 5470-5479.