SEQUENCING AND FUNCTIONAL ANNOTATION OF THE BACILLUS-SUBTILIS GENES IN THE 200 KB RRNB-DNAB REGION

The 200 kb region of the Bacillus subtilis chromosome spanning from 25 5 to 275 degrees on the genetic map was sequenced. The strategy applie d, based on use of yeast artificial chromosomes and multiplex Long Acc urate PCR, proved to be very efficient for sequencing a large bacteria l chromosome area. A total of 193 genes of this part of the chromosome was classified by level of knowledge and biological category of their functions, Five levels of gene function understanding are defined. Th ese are: (i) experimental evidence is available of gene product or bio logical function; (ii) strong homology exists for the putative gene pr oduct with proteins from other organisms; (iii) some indication of the function can be derived from homologies with known proteins; (iv) the gene product can be clustered with hypothetical proteins; (v) no indi cation on the gene function exists. The percentage of detected genes i n each category was: 20, 28, 20, 15 and 17, respectively. In the seque nced region, a high percentage of genes are implicated in transport an d metabolic linking of glycolysis and the citric acid cycle. A functio nal connection of several genes from this region and the genes close t o 140 degrees in the chromosome was also observed.