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Revised genetic linkage map of Bacillus subtilis

Bacillus subtilis is the most widely studied and thoroughly mapped gram-positive microorganism. Two of the driving forces behind the study of the genetics of B. subtilis are the quest to understand the simple differentiation cycle of sporulation and germination that this organism undergoes and the basis for copious enzyme excretion in sporulating organisms. The study of these processes has resulted in the accumulation of vast amounts of knowledge of the physiology and genetics of postexponential bacteria. This is reflected in the abundance of loci that have to do with either this phase of the bacterial life cycle or sporulation and germination. In the several years that have passed since the last compilation of a genetic map for this organism (110, 111) there have been substantial additions to the genetic map. Some 140 loci have been added, making a total of about 500 loci. In addition, there has been refinement of the map in several areas. This has resulted not only from the continuation of classical genetic studies that use deoxyribonucleic acid (DNA)-mediated transformation and tranduction, but also from studies of the molecular cloning of genes of interest in this organism. Several libraries ofB. subtilis DNA have been constructed in bacteriophage X (68) and cosmid (14) and other vectors (131, 243), and these libraries have been a rich source of fragments of the chromosome carrying genes of interest. There have been two important advances in the ways of identifying mutations. The first is the use of integrational vectors carrying cloned B. subtilis DNA (69). These can integrate into the chromosome at the region of homology by Campbell-like mechanism and in so doing place a drug-resistant determinant on the chromosome next to the cloned region. The second is the use of the transposon Tn917, which places a drug resistance marker within mutations it causes. There is now a substantial collection of Tn9J7 insertions generated primarily by Youngman, Losick, and colleagues (331). None of these insertions have been included in the present map, primarily because we did not want to deal with problems of nomenclature for such insertions at this time. Any future revision of the map will have to take them into account. It seems likely that the density of genetic markers on the map as we presently know it will double or triple because of the increase in transposon mutants as well as the identification of new open reading frames from cloned DNA fragments.