Chassis/B.subtilis key parts


B.subtilis strains B. subtilis chassis characterisation B. subtilis specific parts Key Parts and Devices

Bacillus subtilis Key Parts

This page is aimed to highlight some of the key biobricks for B. subtilis that are available on the Registry of Standard Biological Parts.

AmyE Integration Biobricks

IntegrationAmyE.PNG

The AmyE integration bricks (BBa_K143001 and BBa_K143002) are composed of two biobricks that can be added to the 5' and 3' ends of a construct to allow integration into the B. subtilis genome. These biobricks have been successfully used within the constructs BBa_K143079 and BBa_K143082for integration. The possible advantages of integration are within stability and control of copy number. For more information about this biobrick please see [http://2008.igem.org/Team:Imperial_College Imperial College 2008 iGEM project.]

Constitutive Promoters

TestingPromoters.PNG

The pVeg promoter and spoVG ribosome binding site (RBS) Biobrick (BBa_K143053) has been characterised for expression of both GFPmut3b (BBa_E0040) and mRFP1 (BBa_E1010). This data not only shows the parts are both working but once further parts have been characterized provides a basis for comparison. Further promoter and RBS combinations are currently being characterized, in addition to the construction of calibration curves to normalise the units of fluorescence to molecules of fluorescent proteins. For more information about this biobrick please see [http://2008.igem.org/Team:Imperial_College Imperial College 2008 iGEM project.]

Multi-host vector pTG262


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 463
    Illegal XbaI site found at 436
    Illegal PstI site found at 424
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 463
    Illegal NheI site found at 5313
    Illegal PstI site found at 424
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 463
    Illegal BglII site found at 2252
    Illegal BamHI site found at 442
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 463
    Illegal XbaI site found at 436
    Illegal PstI site found at 424
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 463
    Illegal XbaI site found at 436
    Illegal PstI site found at 424
    Illegal AgeI site found at 2641
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2427


The BBa_I742123 plasmid was derived from pTG262 by insertion of a biobrick encoding Red Fluorescent Protein between the unique EcoRI and PstI restriction sites. This eliminated the only native XbaI site present in the plasmid, and since there are no native SpeI sites, the resulting plasmid is a compliant BioBrick vector. New biobricks can be inserted into this vector by replacement of the RFP biobrick, and selection of the white colonies.The parent plasmid pTG262 has a multi host replication origin and replicates in a range of gram positive and gram negative bacteria including E. coli, Bacillus subtilis, and Lactobacillus and Lactococcus spp. We hope that it will also replicate in other Gram negative bacteria. At the time of writing we have successfully transformed E. coli and B. subtilis with this construct, selecting with chloramphenicol (15 and 10 mg/l respectively). For more information about this biobrick please see [http://2007.igem.org/Edinburgh Edinburgh 2007 iGEM project.]