Cloning, sequencing, and disruption of the Bacillus subtilis sigma/sup 28/ gene

Journal Article · Fri Apr 01 04:00:00 EST 1988 · J. Bacteriol.; (United States)

OSTI ID:7164799

Helmann, J D; Marquez, L M; Chamberlin, M J

Bacillus subtilis contains multiple forms of RNA polymerase holoenzyme, distinguished by the presence of different specificity determinants known as sigma factors. The sigma/sup 28/ factor was initially purified as a unique transcriptional activity in vegetatively growing B. subtilis cells. Purification of the sigma/sup 28/ protein has allowed tryptic peptides to be prepared and sequenced. The sequence of one tryptic peptide fragment was used to prepare an oligonucleotide probe specific for the sigma/sup 28/ structural gene, and the gene was isolated from a B. subtilis subgenomic library. The complete nucleotide sequence of the sigma/sup 28/ gene was determined, and the cloned sigma/sup 28/ gene was used to construct a mutant strain which does not express the sigma/sup 28/ protein. This strain also failed to synthesize flagellin protein and grew as long filaments. The predicted sigma/sup 28/ product is a 254-amino-acid polypeptide with a calculated molecular weight of 29,500. The sigma/sup 28/ protein sequence was similar to that of other sequenced sigma factors and to the fibB gene product of Escherichia coli. Since the fibB gene product is a positive regulator of flagellar synthesis in E. coli, it is likely that ..beta../sup 28/ functions to regulate flagellar synthesis in B. subtilis.

• Research Organization: Univ. of California, Berkeley (USA)
• OSTI ID: 7164799
• Journal Information: J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 170:4; ISSN JOBAA
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
550400 -- Genetics
59 BASIC BIOLOGICAL SCIENCES
BACILLUS
BACILLUS SUBTILIS
BACTERIA
CELL DIFFERENTIATION
CLONING
DNA SEQUENCING
DNA-CLONING
ENZYMES
GENE REGULATION
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
RNA POLYMERASES
STRUCTURAL CHEMICAL ANALYSIS
TRANSCRIPTION
TRANSFERASES