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Cited by 231

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Genome Engineering Reveals Large Dispensable Regions in Bacillus subtilis

Westers, H.·2003·Molecular Biology and Evolution

Abstract

Bacterial genomes contain 250 to 500 essential genes, as suggested by single gene disruptions and theoretical considerations. If this view is correct, the remaining nonessential genes of an organism, such as Bacillus subtilis, have been acquired during evolution in its perpetually changing ecological niches. Notably, approximately 47% of the approximately 4,100 genes of B. subtilis belong to paralogous gene families in which several members have overlapping functions. Thus, essential gene functions will outnumber essential genes. To answer the question to what extent the most recently acquired DNA contributes to the life of B. subtilis under standard laboratory growth conditions, we initiated a "reconstruction" of the B. subtilis genome by removing prophages and AT-rich islands. Stepwise deletion of two prophages (SPbeta, PBSX), three prophage-like regions, and the largest operon of B. subtilis (pks) resulted in a genome reduction of 7.7% and elimination of 332 genes. The resulting strain was phenotypically characterized by metabolic flux analysis, proteomics, and specific assays for protein secretion, competence development, sporulation, and cell motility. We show that genome engineering is a feasible strategy for functional analysis of large gene clusters, and that removal of dispensable genomic regions may pave the way toward an optimized Bacillus cell factory.

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Metadata

Authors
Westers, H.
Venue
Molecular Biology and Evolution
Published
2003-08-29
Volume
20
Issue
12
Pages
2076-2090
ISSN
  • 0737-4038 (ISSN-L)
  • 1537-1719
OpenAlex
W2124703740
Cited by
231
References
1
FWCI
3.68
Type
journal article
Language
en
Open access
closed

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