Mapping a cloned gene under sporulation control by inserttion of a drug resistance marker into the Bacillus subtilis chromosome.
A segment of Bacillus subtilis deoxyribonucleic acid (DNA) previously cloned in Escherichia coli contains a gene (the 0.4-kilobase [kb] gene) whose transcription is activated at an early stage of spore development. To map the genetic location of the 0.4-kb gene, we constructed a hybrid plasmid that inserts a chloramphenicol resistance determinant into the B. subtilis chromosome by recombination at a site of homology between cloned B. subtilis DNA and the chromosome. This hybrid plasmid (p1949-2) was constructed from the E. coli plasmid pMB9, the B. sultilis chloramphenicol resistance plasmid pCM194 (whose replication function was inactivated), and B. subtilis DNA from the vicinity of the 0.4-kb gene. Transformation of B. subtilis cells to drug resistance by p1949-2 was dependent upon the B. subtilis RecE+ phenotype and resulted in specific and predictable changes in the pattern of endonuclease restriction sites in the 0.4-kb gene region of the chromosome. Chloramphenicol resistance in cells transformed by p1949-2 was mapped to the purA-cysA region of the B. subtilis chromosome, a region. In addition, DNA adjacent to the 0.4-kb gene was shown to contain the wild-type allele of genetic marker (tms-26) from that region.
D001412:Bacillus subtilis / Q000235:genetics* / Q000502:physiology; D002701:Chloramphenicol / Q000494:pharmacology; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D005796:Genes; D005819:Genetic Markers; D010957:Plasmids; D013171:Spores, Bacterial / Q000235:genetics; D014169:Transformation, Bacterial
D005819:Genetic Markers; D002701:Chloramphenicol