Characterization and structure of the cellulase gene of Bacillus subtilis BSE616.
The Bacillus subtilis carboxymethyl cellulase (CMCase) gene originally cloned on a 3.2-kb PstI DNA fragment has been localized in a 1.5-kb Sau3AI fragment by a series of subclonings into plasmid pUC19. During the process the promoter region and Shine-Dalgarno (SD) sequence were deleted, but the 1.5-kb insert was shown to direct the synthesis of CMCase in scherichia coli to a high level, probably with the aid of lac promoter. We analyzed the complete nucleotide sequence of the CMCase gene. The CMCase gene is 1500-bp long, encoding a polypeptide of 499 amino acids and a stop codon. The putative "-35" region (TAGACA), "-10" region (TACAAT), and ribosome binding site (RBS) (AAGGAGG) have also been identified in the 5' flanking region. Comparison of the nucleotide sequence to three other published endo-beta-1,4-glucanase genes of B. subtilis strains shows that these sequences share very strong homology. It seems that the cellulase genes have been derived from a common ancestor by spontaneous mutation. The probability of carboxy-terminal processing of the CMCase protein is also discussed.
Department of Biological Science and Engineering, Korea Advanced Institute of Science and Technology, Seoul.
S H
SH
Park
H K
HK
Kim
M Y
MY
Pack
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D002480:Cellulase; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D006026:Glycoside Hydrolases / Q000235:genetics*; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D004269:DNA, Bacterial; D006026:Glycoside Hydrolases; D002480:Cellulase; C019826:carboxymethylcellulase