Difference between revisions of "Part:BBa K2797002"

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<partinfo>BBa_K2797002 short</partinfo>
 
<partinfo>BBa_K2797002 short</partinfo>
 
 
This composite part confers resistance to streptomycin. The part contains the constitutive BBa_J23119 promoter, the strongest promoter of the J23100 through to J23119 promoter family
 
This composite part confers resistance to streptomycin. The part contains the constitutive BBa_J23119 promoter, the strongest promoter of the J23100 through to J23119 promoter family
  

Revision as of 14:39, 2 October 2018


Composite part for streptomycin resistance This composite part confers resistance to streptomycin. The part contains the constitutive BBa_J23119 promoter, the strongest promoter of the J23100 through to J23119 promoter family

Part Characterisation

T--NEWCASTLE--STREPTOMYCIN ECOLISMR.jpeg

Figure 1 E. coli strain DH5α transformed with streptomycin resistance part BBa_K2797002 growing on LB agar containing streptomycin at a concentration of 50 μg/ml.

T--NEWCASTLE--STREPTOMYCIN ECOLI.jpeg

Figure 2 Untransformed E. coli strain DH5α unable to grow on LB agar containing streptomycin at a concentration of 50 μg/ml.

T--NEWCASTLE--SMRCMA.jpeg

Figure 3 E. coli strain DH5α transformed with streptomycin resistance part BBa_K2797002 growing on LB agar containing chloramphenicol at a concentration of 25 μg/ml due to resistance gene in pSB1C3 vector.

T--NEWCASTLE--Strep.jpeg

Figure 4 Optical density at 600nm of three replicate wells containing LB and incrementally increasing streptomycin concentrations inoculated with E. coli strain DH5α untransformed and transformed with part BBa_K2797002 (SmRp) during incubation at 37 C over 24 hours. Control wells contained untransformed E. coli DH5α and an antibiotic concentration of zero. Error bars show standard error mean.

Usage and Biology

A selection of resistance genes are required for selection of transformants when inserting foreign DNA into a chassis organism. Newcastle iGEM 2018 focused on developing a new chassis organism capable of colonising plant roots. The selected organism, Pseudomonas fluorescens DSM 25356 was found to be resistant to a range of commonly used antibiotics. Streptomycin was one of two antibiotics P. fluorescens was found not to be resistant to. The team therefore decided to design a streptomycin resistance part that could in future be used for selection of transformed P. fluorescens.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 861
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 709
  • 1000
    COMPATIBLE WITH RFC[1000]