In collaboration with the Department of Olli Lassila at Turku University in Finnland we are building filter and glas slide gene arrays which could be used to quantitate gene expression profiles in wild-type and mutant DT40 cells. This bursal EST sequencing project will continue for the next three years. To avoid redundant sequencing of highly expressed genes we have successfully normalized the bursal cDNA library by hybridization with already sequenced ESTs. This library is currently orderedin the German Genome Resource Center. Using the normalized library and the library enriched for full-length clones we are planning to add new sequences at a rate of 1000 sequences per month for the next three years.
All ESTs are freely available to all researchers.
We are producing other generic resources for genetic studies in DT40. In the past, six drug resistance markers were available, but for analysis of complex pathways a higher number of mutations are needed. We therefore constructed new markers flanked by loxP sites which are recognized by the CRE-recombinase. These markers allow the generation of unlimited number of mutations in DT40 through the reuse of the markers after CRE-mediated excision.