Promoters from a cold-adapted bacterium: definition of a consensus motif and molecular characterization of UP regulative elements.
Although low-temperature tolerant micro-organisms were discovered long ago, limited information is still available on the transcription machinery in cold-adapted bacteria. This knowledge represents a necessary background for the investigation of the adaptation of gene-expression mechanisms at low temperatures. The recent development of a shuttle genetic system for the transformation of the cold-adapted Gram-negative bacterium Pseudoalteromonas haloplanktis strain TAC125 has made possible the isolation of the psychrophilic promoters described in this paper. TAC125 genomic DNA fragments were cloned in the shuttle vector and the promoter-containing recombinant clones were selected for their ability to express a promoter-less lacZ gene. The nucleotide sequence of several selected inserts and the transcription start points of the transcribed m-RNAs were determined. A promoter consensus sequence for Pseudoalteromonas haloplanktis TAC125 was proposed on the basis of a sequence comparison between the various active promoters. Furthermore, the identification and the functional characterization of two UP elements from this cold-adapted bacterium are also reported.
Duilio
Angela
A
Dipartimento di Chimica Organica e Biochimica, Università di Napoli "Federico II", Complesso Universitario Monte Sant'Angelo, via Cynthia, 80126, Naples, Italy. anduilio@unina.it
Madonna
Stefania
S
Tutino
Maria Luisa
ML
Pirozzi
Marinella
M
Sannia
Giovanni
G
Marino
Gennaro
G
D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D001483:Base Sequence; D003080:Cold Temperature; D016384:Consensus Sequence / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D031161:Pseudoalteromonas / Q000235:genetics* / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*