Disruption of the ADGF multigene family by homologous recombination. T. Dolezal ^1,2,3, M. Gazi ^1,2,3, M. Zurovec ^1,2, P.J. Bryant ¹. 1) Dept DBC, Univ California, Irvine, Irvine, CA; 2) Institute of Entomology and University of South Bohemia, Ceske Budejovice, Czech Republic; 3) Authors contributed equally to this work.

   We have recently identified a novel family of six Adenosine Deaminase-related Growth Factors (ADGF) in Drosophila. These secreted proteins stimulate proliferation of various cell lines by depleting extracellular adenosine, which is otherwise cytotoxic (Zurovec et al., 2002). In order to investigate the functions of these factors in vivo we are using targeted mutagenesis by homologous recombination (Rong and Golic, 2000) to obtain mutations in five of the six family members. For this procedure we are taking advantage of the fact that five of the family members are encoded by a three-gene cluster and a two-gene cluster. We designed two constructs for ends-in targeting to introduce loss of function mutations in all five genes as well as different combinations of multiple mutations in just two independent crossing schemes. Loss-of-function mutations in ADGF-A and ADGF-C show a larval and pupal lethal phenotype, respectively, consistent with a requirement for the gene products for larval growth and survival.
   Rong, Y.S. and Golic K.G. (2000). Gene Targeting by homologous recombination in Drosophila. Science 288:2013.
   Zurovec, M. et al. (2002). Adenosine deaminase-related growth factors stimulate cell proliferation in Drosophila by depleting extracellular adenosine. Proc Natl Acad Sci U S A. 99:4403.
   
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