Difference between revisions of "Part:BBa K2797002"

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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
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===Usage and Biology===
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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, <I>Pseudomonas fluorescens</I> DSM 25356 was found to be resistant to a range of commonly used antibiotics. Streptomycin was one of two antibiotics <I>P. fluorescens</I> 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 <I>P. fluorescens</I>.
  
 
===Part Characterisation===
 
===Part Characterisation===
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'''Figure 4''' Optical density at 600nm of three replicate wells containing LB and incrementally increasing streptomycin concentrations inoculated with <I>E. coli</I> strain DH5α untransformed and transformed with part BBa_K2797002 (SmR<I>p</i>) during incubation at 37 C over 24 hours. Control wells contained untransformed <I>E. coli</I> DH5α and an antibiotic concentration of zero. Error bars show standard error mean.
 
'''Figure 4''' Optical density at 600nm of three replicate wells containing LB and incrementally increasing streptomycin concentrations inoculated with <I>E. coli</I> strain DH5α untransformed and transformed with part BBa_K2797002 (SmR<I>p</i>) during incubation at 37 C over 24 hours. Control wells contained untransformed <I>E. coli</I> 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, <I>Pseudomonas fluorescens</I> DSM 25356 was found to be resistant to a range of commonly used antibiotics. Streptomycin was one of two antibiotics <I>P. fluorescens</I> 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 <I>P. fluorescens</I>.
 
  
 
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Revision as of 14:41, 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

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.

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.

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]