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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.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

INSTITUT PASTEUR
Address
25,Rue Du Docteur Roux 28
75724 Paris
France

Participants (14)

AGOWA - Gesellschaft fur Molekularbiologische Technologie mbH
Germany
Address
185,Glienicker Weg 185
12489 Berlin
GATC - Gesellschaft fur Analyse-Technik und Consulting mbH
Germany
Address
23,Fritz-arnold-str. 23
78467 Konstanz
Genotype GmbH, Molekularbiologische und biotechnologische Forschung
Germany
Address
39,Angelhofweg
Wilhelmsfeld
Institut National de la Recherche Agronomique
France
Address
Domaine De Vilvert
78352 Jouy-en-josas
International Centre for Genetic Engineering and Biotechnology
Italy
Address
Padriciano 99
34012 Trieste
QIAGEN GMBH
Germany
Address
4,Max-volmer-strasse 4
40724 Hilden
SCIENTIFIC RESEARCH AND DEVELOPMENT GMBH
Germany
Address
43,Oberurseler Strasse 43
61440 Oberursel
THE PROVOST, FELLOWS AND SCHOLARS OF THE COLLEGE OF THE HOLY AND UNDIVIDED TRINITY OF QUEEN ELIZABETH NEAR DUBLIN HEREINAFTER TRINITY COLLEGE DUBLIN
Ireland
Address
Smurfit Institute,trinity College
2 Dublin
UNIVERSITA DEGLI STUDI DI PAVIA
Italy
Address
Via Ferrata 1
27100 Pavia
UNIVERSITE CATHOLIQUE DE LOUVAIN
Belgium
Address
2/20,Place Croix Du Sud 2/20
1348 Louvain-la-neuve
UNIVERSITY OF GRONINGEN
Netherlands
Address
30,Kerklaan 30
9750 AA Haren Gn
UNIVERSITY OF NEWCASTLE UPON TYNE
United Kingdom
Address
Framlington Place
NE2 4HH Newcastle - Upon Tyne
Università degli Studi di Padova
Italy
Address
Via Trieste 75
35121 Padova
Université de Lausanne
Switzerland
Address
19,Rue Cesar-roux
1005 Lausanne