Reporter

Part:BBa_J33206

Designed by: Chris French   Group: iGEM2006_Edinburgh   (2006-10-26)


Bacillus subtilis ars promoter and arsR gene plus E. coli lacZ

This part includes the arsenate-responsive ars promoter from Bacillus subtilis (Sato, T., and Kobayashi, Y. 1998. J. Bacteriol. 180, 1655-1661), together with the arsR gene encoding the negatively autoregulated repressor ArsR, plus the Escherichia coli lacZ' gene encoding the N-terminal 137 amino acid residues of LacZ, sufficient to complement the chromosomal lacZ-delta-M15 mutation carried by many common lab strains of E. coli. This construct is designed to give LacZ' expression in the presence of arsenic as arsenate or arsenite anion. In previous experiments using other reporter genes such as xylE (available as biobrick BBa_J33204), we have found that this promoter gives a more rapid and stronger response than the E. coli chromosomal ars promoter (available as biobrick BBa_J33201, and fused to lacZ' as BBa_J33203), but with higher background activity in the absence of arsenate. Our experiments with this particular construct showed no obvious inducibility of a pH response due to a high background expression level in the absence of arsenate; however, ONPG assays have not yet been done. This construct does not include a terminator, so another reporter gene could be added at the 3' end for further experiments.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 454


[edit]
Categories
//chassis/prokaryote/bsubtilis
Parameters
None