Difference between revisions of "Part:BBa K2797002"
(→Characterisation) |
|||
| (7 intermediate revisions by 3 users not shown) | |||
| Line 6: | Line 6: | ||
===Usage and Biology=== | ===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, < | + | 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, <i>Pseudomonas</i> sp. strain CT364 was found to be resistant to a range of commonly used antibiotics. Streptomycin was one of two antibiotics <i>Pseudomonas</i> sp. was found not to be resistant to. The team has deposited a new streptomycin resistance cassette. |
===Characterisation=== | ===Characterisation=== | ||
| Line 23: | Line 23: | ||
[[File:T--NEWCASTLE--Strep4.jpeg|900px|]] | [[File:T--NEWCASTLE--Strep4.jpeg|900px|]] | ||
| − | '''Figure 4''' <I>E. coli</I> DH5α with or without the BBa_K2797002 part in pSB1C3 were grown in LB | + | '''Figure 4''' <I>E. coli</I> DH5α with or without the BBa_K2797002 (SmR<I>p</I>) part in pSB1C3 were grown in LB broth containing streptomycin at varying concentrations. Cells were grown in 96-well plate format in 200 μl volumes at 37 C over 24 hours. (n=3 replicates, error bars are standard error of the mean). |
<!-- --> | <!-- --> | ||
Latest revision as of 23:22, 17 October 2018
Composite part for streptomycin resistance
This composite part confers resistance to streptomycin. The part contains the constitutive promoter BBa_J23119 from the Anderson collection, BBa_B0034 ribosome binding site, BBa_K125520 coding sequence conferring resistance to streptomycin and BBa_K2797001 terminator.
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 sp. strain CT364 was found to be resistant to a range of commonly used antibiotics. Streptomycin was one of two antibiotics Pseudomonas sp. was found not to be resistant to. The team has deposited a new streptomycin resistance cassette.
Characterisation
Figure 1 E. coli strain DH5α transformed with streptomycin resistance part BBa_K2797002 plated on LB agar containing streptomycin (50 μg/ml).
Figure 2 E. coli strain DH5α plated on LB agar containing streptomycin (50 μg/ml).
Figure 3 E. coli strain DH5α carrying the streptomycin resistance part BBa_K2797002 plated on LB agar containing chloramphenicol (25 μg/ml). The part is in the pSB1C3 backbone conferring resistance to chloramphenicol.
Figure 4 E. coli DH5α with or without the BBa_K2797002 (SmRp) part in pSB1C3 were grown in LB broth containing streptomycin at varying concentrations. Cells were grown in 96-well plate format in 200 μl volumes at 37 C over 24 hours. (n=3 replicates, error bars are standard error of the mean).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 861 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 709
- 1000COMPATIBLE WITH RFC[1000]