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Sequencing of the Bacillus subtilis genome

Objective



Bacillus subtilis is a model organism for Gram-positive bacteria including pathogens, e.g. Staphylococcus, Streptococcus, Corynebacterium, Clostridium and Listeria. It is also an important industrial organism. The determination of the sequence of its genome is therefore an important objective. An international agreement has been reached allowing the allocation of regions of the chromosome among the groups of the European and Japanese Bacillus subtilis genome sequencing networks, also including two biotechnology companies (Genencor International and Novo Nordisk). The European Bacillus subtilis genome sequencing project has been supported in the framework of the current Biotechnology contracts BIO2-CT93-0272 and BIO2-CT942011 which will end in December 1996. A relational database containing more than 2000 kbp of non-redundant Bacillus subtilis DNA sequence (1050 kbp has been contributed by the European network), representing about 60 % of the total genome, has been constructed at the Institut Pasteur. During the last two years the results of this European project have been presented at 13 international meetings, and have been published in several international journals, in particular in a special issue of the journal Microbiology (1995, 141: 249-350). A major difficulty encountered during the project stems from the unavailability of a simple cloning strategy for the construction of gene banks representative of the entire Bacillus subtilis chromosome.
This is, at least in part, due to the generally high level of expression of Bacillus subtilis genes observed in Escherichia coli, leading to toxic effects in this commonly used cloning host. However, participating groups have employed a variety of strategies to overcome this limitation, including cloning into a range of plasmid, lambda and YAC vectors, genome walking experiments and in-vitro amplification using long-range PCR and inverse PCR techniques. Bacillus subtilis is also a major source of industrial enzymes. The world market (estimated at 400 million dollars in 1993) is dominated by the two companies mentioned above (Novo Nordisk and Genencor). An industrial platform has been set up to facilitate contacts between project participants and European biotechnology companies. The following companies have agreed in principle to this mode of cooperation: Novo Nordisk (Denmark, USA), Genencor (Finland, USA) and its subsidiary Gist-brocades (The Netherlands), F. Hoffmann La Roche AG (Switzerland), Eniricerche (Italy), Puratos (Belgium). Thirteen laboratories and five companies, which have a solid experience in large-scale sequencing, agreed to participate in the present proposal. Its objective is the determination of the remaining sequence (estimated at 1800 kbp), leading to the completion of the entire Bacillus subtilis genome sequence within one year.

Call for proposal

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Coordinator

INSTITUT PASTEUR
EU contribution
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75724 PARIS
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Participants (14)