Difference between revisions of "Part:BBa K2797002"

Revision as of 14:42, 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 range of antibiotic 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

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.

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

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.

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