Systematic function analysis of Bacillus subtilis genes

• Research unit: EU RFP
• Project number: 95.0267

Key words (English)

Phage resistance and bacterial competence; Bacillus subtilis; gene function analysis; mutant
construction; mutant screening

Alternative project number (English)

EU project number: BIO4CT950278

Research programs (English)

EU-programme: 4. Frame Research Programme - 4.1 Biotechnology

Short description (English)

See abstract

Further information (English)

Full name of research-institution/enterprise:
Université de Lausanne
Institut de Génétique et de Biologie microbiennes

Abstract (English)

Sequencing of the 'entire Bacillus subtilis genome by a Euro-Japanese consortium (Kunst et al., 1997) was followed by a new Euro-Japanese project 'Systematic Function Analysis of B. subtilis Genes' which essentially consisted in the inactivation of relevant genes and the characterization of thus obtained mutants.
Mutant construction. The hisA-gerB (306 0-315 0) and cotT-rapA (109 0-112 0) regions of the chromosome were assigned to IGBM. All relevant orphan genes were inactivated. Two genes are still under examination. 71 knock-out mutants were constructed and verified by PCR-tests. For all 71 strains, beta-galactosidase expression tests were performed in rich and minimal media in different growth phases. Fourteen genes showed no expression at all. One strain failed to grow in minimal medium, while two others grew at a markedly reduced rate in both media. Au data on strain construction and beta-gal expression have been introduced in the Micado Data Base.
(http://locus.jouy.infra.fr/cgi-bin/genmic/madbase/progs/madbase.operl)
Mutant screening. Over 1200 mutants have been obtained by the European participants of the consortium. All of them have been screened for growth, modified phage resistance phenotype and altered transformability. One strain was resistant to phage rho 11 and devoid of the poly(glucosyl-N-acetyl-galactosamine-phosphate) teichoic acid. Three strains showed reduced competence (1 to 30% of wild type).
Further analysis. Growth tests with mutants in genes of the yjm-operon confirmed its involvement in hexuronate catabolism, in agreement with sequence homology. Growth tests with nag mutants confirmed that nagA is a N-acetyl-glucosamine deacetylase, but the actual deaminase appeared to be ybfT, and not nagB. Biochemical and physiological characterization of the yvoB (ptsK), yvoC (gerF) and cccB gene products led to full articles (see references).
References
Reizer 3. et al. (1998). A novel protein kinase that controls carbon catabolite repression in bacteria. Microbiology 27, 1157-69.
Robinson C. et al. (1998). The product of the yvoC (gerF) gene of Bacillus subtilis is requil~d for spore germination. Microbiology 144, 3105-3109.
Bengtsson 3. et al. (1999). Subunit II of Bacillus subtilis cytochrome C oxidase is a lipoprotein. 3. Bacteriol. 131, 685-8.
Bengtsson 3. et al. (submitted). Bacillus subtilis contains two small c-type cytochromes with homologous heme-domains but different types of membrane-anchors.

References in databases (English)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 95.0267