MAPPING THE TRYPANOSOMA-CRUZI GENOME - ANALYSES OF REPRESENTATIVE COSMID LIBRARIES

In order to generate contiguous cosmid coverage of the genome of the p rotozoan parasite Trypanosoma cruzi for large-scale sequence analysis, a cosmid library of 36 864 individual, primary clones was generated. Total genomic DNA of the reference strain CL Brener was fragmented bot h by partial digestion with MboI and by physical shearing. For cloning , a modified cosmid vector was used that simplifies analyses such as r estriction mapping. The library's representation is about 25 genome eq uivalents, assuming a size of 55 Mb per haploid genome. No chimerism o f inserts in the clones could be detected. The colinearity between cos mid inserts and genomic DNA was verified Also, hybridizations to the g el-separated karyotype of the organism were carried out as a quality c heck. Gridded onto two nylon filters, the library was analyzed with a variety of probes. Apart from being used for combined physical and tra nscriptional mapping of the genome, library filters and clones are als o available to interested parties.