ESTABLISHMENT OF CHROMOSOME MAPPING OF WHOLE BAC CLONES IN SCHISTOSOMA MANSONI.

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This project has promoted objectives that develops techniques of chromosome mapping of BAC clones and establishes address of all the clones, as a part of genome project of Schistosoma mansoni. We have done following items during the last four years of the project.(1)We improved adequate techniques for chromosome preparation, labeling of BAC clone DNA, denaturation of chromosome DNA, and measuring location of signal on chromosome. These improvements allowed us to establish FISH (fluorescence in situ hybridization) for chromosome mapping. The techniques were available for filarial chromosomes as well.(2)Because chromosomes 5,6, and 7 of Schistosoma mansoni are little in size and similar in shape, they are very difficult to be identified by only their shape if you are not familiarized to identifying them. Therefore, we produced specific painting probes to each chromosome in order to improve the objective technique of chromosome identification.(3)Establishment of chromosome mapping of BAC clones allowed us to localize some specific genes on chromosomes, for instance adenylosuccinate lyase and HOX genes.(4)To supplement for FISH technique, PRIS(primed in situ) labeling technique, that makes PCR amplification of DNA on chromosomes, was applied to chromosomes of Schistosoma mansoni.(5)Techniques that have been established in the project were also applied to experiments of gene mapping and chromosome differentiation in primates.The first aim of the project that makes chromosome mapping of 20000 BAC clones was not able to attain yet. However, it will be possible according to collaboration of other institutions in the world. Our techniques are going to handover to other project teams of parasite genome project, so that outcomes produced in the project will contribute for development of the parasite genome project on the move in the world.