Difference between revisions of "Part:BBa K2797002"

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

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]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <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 promoter BBa_J23119 from the Anderson collection, BBa_B0034 ribosome binding site, BBa_K125520 coding sequence conferring resistance to streptomycin and BBa_K2797001 terminator.
-===Part Characterisation===
+===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, <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===
 [[File:T--NEWCASTLE--STREPTOMYCIN ECOLISMR.jpeg|500px|]]
-'''CFPS Expression of ''J23100-deGFP'' and ''J23100-sfGFP'' Constructs:''' Time course for increase in fluorescence intensity of CFPS systems expressing GFP constructs over time. Each data point is an average of triplicate results, and error bars show +/- standard error. CFPS system with no DNA (Red) was used as the negative control.
+'''Figure 1''' <I> E. coli </I> strain DH5α transformed with streptomycin resistance part BBa_K2797002 plated on LB agar containing streptomycin (50 μg/ml).
+[[File:T--NEWCASTLE--STREPTOMYCIN ECOLI.jpeg|500px|]]
-<!-- Add more about the biology of this part here
+'''Figure 2''' <I> E. coli </I> strain DH5α plated on LB agar containing streptomycin (50 μg/ml).
-===Usage and Biology===
+[[File:T--NEWCASTLE--SMRCMA.jpeg|500px|]]
+'''Figure 3''' <I> E. coli </I> 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.
+[[File:T--NEWCASTLE--Strep4.jpeg|900px|]]
+'''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).
 <!-- -->