Open AccessThe xylA promoter of Bacillus megaterium mediates constitutive gene expression in Escherichia coli
Author(s) -
Knobloch Denise,
Clemens Andre,
Ostermann Kai,
Rödel Gerhard
Publication year - 2011
Publication title -
engineering in life sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.547
H-Index - 57
eISSN - 1618-2863
pISSN - 1618-0240
DOI - 10.1002/elsc.201000225
Subject(s) - bacillus megaterium , shuttle vector , biology , escherichia coli , microbiology and biotechnology , heterologous expression , promoter , gene , open reading frame , gene expression , heterologous , recombinant dna , vector (molecular biology) , bacteria , genetics , peptide sequence
In the present study, we demonstrate that the Escherichia coli–Bacillus megaterium shuttle vector pHIS1522 can be used as a versatile expression vector. Recombinant genes under the control of the xylA promoter are constitutively expressed at a high level in E. coli strains, whereas their expression is strongly induced by the addition of xylose in B. megaterium . The utilization of D ‐xylose is known to be dependent on the xylAB genes in a number of bacteria. For B. megaterium a XylA‐based expression system was established that allows tightly regulated and highly efficient heterologous gene expression. The open reading frame (ORF) of the fluorescent protein turboRFP was cloned under the control of the xylA promoter of B. megaterium in the shuttle vector pHIS1522 . Unexpectedly, tRFP expression was not only observed in B. megaterium, but also in E. coli . Based on fluorescence measurements and Western blot analysis, expression was comparable or slightly higher compared with the commonly used pET vectors. Therefore, pHIS1522 can be used as a versatile expression vector in both, B. megaterium and E. coli.