Coding

Part:BBa_K1021003

Designed by: Swati Sureka   Group: iGEM13_Cornell   (2013-08-30)


bar (bialaphos resistance)

The bialaphos resistance (bar) gene confers resistance to bialaphos and phosphinothricin.

Usage and Biology

The common fungus transformation system of entomopathogen Metarhizium anisopliae was based on resistance to the herbicide glufosinate ammonium (phosphinothricin), conferred by the bar gene.

Team NYMU 2016 characterized the bar marker gene, a twin of this part, using the bar expression cassette on vector pBARGPE1 with our chassis organism M.anisopliae.

The transformants carrying the bar gene can grow on the PDA containing glufosinate ammonium.

Figure1. The emerged colonies on regeneration medium
Figure2. Transformants on selective medium



We did the glufosinate ammonium inhibition test using three different concentrations (0, 160, 1000 ug/mL) with bar transformants and wildtype as a control, the result is showed below.

The initial radius were all 4mm. The mycelium length was measured on day 4, 5 and 6 after starting M.anisopliae conidia cultivation on solid media. The mycelium length was measured on the same line on each plate.

Bar a.png
Bar-6.png


  • The higher glufosinate ammonium concentration, the more sparse the mycelium grows outward.


Data:

WT-0, 160, 1000 ug/mL: Wild type strains grew on the media containing 0, 160, 1000 ug/mL glufosinate ammonium.


Bar-0, 160, 1000 ug/mL: Bar transformants grew on the media containing 0, 160, 1000 ug/mL glufosinate ammonium.

The mycelium length on 0, 4, 5, 6 days


Bar 1.png





Bar2.png



These results showed that glufosinate ammonium affect the growth of the mycelium, but not as effectively as expected. It may be caused by the degradation of this herbicide due to the long store.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 545
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 54
    Illegal NgoMIV site found at 205
    Illegal NgoMIV site found at 451
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//cds/selectionmarker/antibioticresistance
Parameters
None