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Zawartość zarchiwizowana w dniu 2024-04-30

Systematic function analysis of Bacillus subtilis genes

Cel



The primary objective of the proposal is to analyse genes, uncovered by sequencing of the B. subtilis genome in Europe, which encode products that show either no similarities with known proteins or carry only short domains or "signatures" which are not sufficient for identification of their function. The target genes will be assigned to large categories of known cell functions, defined here as: (i) Metabolism of small molecules and inorganics; (ii) Macromolecule metabolism; (iii) Cell structures and mobility; (iv) Stress and stationary phase; (v) Cell processes. About 1000 target genes will be analysed, which is over 20% of the estimated 4 200 B. subtilis genes.
The assignment will be based mainly on an extensive phenotype analysis of the mutants in target genes and a biochemical analysis of transcripts and proteins, as well as the whole cell composition. Two research consortia will be established to carry out these analyses, the resource consortium and the function-oriented consortium, associating 19 laboratories. The activities of each consortium will be organized in several nodes. The resource consortium comprises four nodes, (i) mutant construction and distribution; (ii) RNA analysis; (iii) protein analysis and cell composition; (iv) database development. A standard procedure will be used to construct the mutants, allowing to follow at the same time the regulation of the target gene and to regulate the genes downstream in the operon. The mutants will be stored in a central collection and distributed throughout the function- oriented consortium. RNA analysis will allow to generate a transcription map of at least 750 kb in several long contigs, and high through-put methods will be developed and tested to extend it further. Protein analysis will be carried out by two dimensional gel electrophoresis in over 700 mutants, aiming to detect regulator mutants and to progress towards a complete 2D protein data base. Finally, a user friendly, easily accessible data base will be developed and used to store the information generated within the programme and to correlate it with the sequence databases.
The nodes of the function-oriented consortium correspond to the five categories of cell functions specified above. To assign target genes to these categories the analysis will be carried out at three levels. At the first, which aims to a tentative assignment, the mutants will be submitted to phenotype tests that have a sufficiently high through-put to allow analysis of all mutants by each participating laboratory. Between 10 and 100 tests will be carried out in each laboratory, depending on the category of cell function. At the second level, aiming to confirm the gene assignment and to initiate the studies of the role of the target genes, the mutants will be submitted to more elaborate tests, having a through-put that allows testing of 5-10% of all mutants in each laboratory. The third level analysis aims to identify the specific role of a small number of genes that appear of a particular interest. Among these will be genes of bio-technological interest and genes that have homologues in many organisms, and specifically yeast S. cerevisiae.

Dziedzina nauki

Program(-y)

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System finansowania

CSC - Cost-sharing contracts

Koordynator

Institut National de la Recherche Agronomique
Wkład UE
Brak danych
Adres
Domaine de Vilvert
78352 Jouy-en-Josas
Francja

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Koszt całkowity
Brak danych

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Ostatnia aktualizacja: 22 Października 1998

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Permalink: https://cordis.europa.eu/project/id/BIO4950278/pl

European Union, 2024