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Transformation of Nematodes With the Helios Gene Gun

Peter Jackstadt1, Horst Zahner2 and Gerd Hobom1, 1 Institut fr Mikrobiologie und Molekularbiologie der Justus-Liebig Universitt Gieen, Frankfurter Str. 107, D-35392, Gieen, 2 Institut fr Parasitologie der Justus-Liebig-Universitt Gieen, Rudolf-Buchheim-Str. 2, D-35392 Gieen


Introduction
The Biolistic DNA transfer technique was first established for plant cells and plant tissues (Sandford et al., 1987; Jefferson et al., 1987; Bruce et al., 1989; Armaleo et al., 1990). More recent applications have been DNA transfer experiments aiming at DNA vaccination of animals (genetic immunization; Sundaram et al., 1996), for example a bombardment of mice to generate protective antibodies, instead of the DNA injection used for this purpose in the past.

Here we report on the potential to use this convenient method for transfection of nematodes, not only for the free-living soil nematode Caenorhabditis elegans, the well known model organism in its class, but also for the cotton rat filaria Litomosoides sigmodontis, which serves as model for human pathogenic filariae (Zahner, Hobom and Stirm, 1995), such as Wuchereria bancrofti or Brugia spp. These parasitic species cause a severe lymphatic disease called filariasis (elephantiasis), from which more than 100 million human beings suffer worldwide.


Methods
PREPARATION OF ADULT HERMAPHRODITE C. ELEGANS AND COTTON RAT PARASITE L. SIGMODONTIS FOR MICROPARTICLE BOMBARDMENT
C. elegans is cultivated on NGM agar plates (Brenner, 1974). Wild-type worms (strain N2 var. Bristol) are fed on Escherichia coli OP-50 (exogenous uracil dependent mutant) seeded on those pla
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