Abstract
To establish a constitutive, high-efficiency expression system for Bacillus pumilus (B.P), we cloned random chromosomal DNA into promoter probe shuttle vector ECE7 and selected for strong promoter activity by chloramphenicol resistance of transformed B. pumilus cells. The nucleotide sequences of nine chromosomal fragments were determined. These DNA fragments range from 300 to 2200 bp in size. The transcription strength of these promoters was estimated by determination of CAT enzyme production in both E. coli and B. pumilus. Transcription start (TS) sites of the cat mRNA were located by primer extension by using total RNA. Preliminary analysis showed that three of the promoter sequences contain −35 and −10 regions like E. coli RNA polymerase ς70 and B. subtilisς43 consensus sequences. One is similar to B. subtilisς29, the other two have no conserved sequences like any of the typical consensus sequences of the known sigma factors so far. To estimate the feasibility of the utilization of these promoters, one promoter fragment was subcloned and used to drive the expression of green fluorescent protein (GFP) in B. pumilus cells. This is the first report of B. pumilus promoters randomly cloning from total DNA and molecular analysis of their consensus sequences.
Similar content being viewed by others
Author information
Authors and Affiliations
Additional information
Received: 27 December 2000 / Accepted: 6 February 2001
Rights and permissions
About this article
Cite this article
Cao, Q., Qu, Z., Wan, Y. et al. Cloning, Molecular Characterization, and Application of Rice Epiphytic Bacillus pumilus Promoter Fragments. Curr Microbiol 43, 244–248 (2001). https://doi.org/10.1007/s002840010295
Issue Date:
DOI: https://doi.org/10.1007/s002840010295