Difference between revisions of "Part:BBa K2999003"

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Biofilm is a complex bacterial community embedded in the extracellular matrix composed of proteins, extracellular DNA (eDNA) and extracellular polysaccharides. Pseudomonas aeruginosa PAO1 has two types of biofilm, Psl and Pel.
 
Biofilm is a complex bacterial community embedded in the extracellular matrix composed of proteins, extracellular DNA (eDNA) and extracellular polysaccharides. Pseudomonas aeruginosa PAO1 has two types of biofilm, Psl and Pel.
 
This part is mainly used for encoding polysaccharide hydrolases PslG and PelA, and destroying the structure of biofilm.
 
This part is mainly used for encoding polysaccharide hydrolases PslG and PelA, and destroying the structure of biofilm.
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===Usage and Biology===
 
===Usage and Biology===
 
YAU-China expressed the biofilm-damaged system in 2019, using extracts from each part of the system for experiments.
 
YAU-China expressed the biofilm-damaged system in 2019, using extracts from each part of the system for experiments.
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The control group (PBS treated biofilm), PelA treated biofilm, PslG treated biofilm, PelA and PslG treated biofilm together. The biofilm was added to the PV plate and pretreated for 24 hours. The biofilm was washed out with absolute ethanol and detected by enzyme labeling instrument.
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After overnight induction of the control strain, two strains expressing single enzyme and two strains co-expressing two enzymes, four strains were collected by centrifugation and suspended in PBS buffer of equal volume. The supernatant was absorbed and added into the biofilm of Pseudomonas aeruginosa cultured on the pre-PVC plate. After incubation for a period of time, the PVC plate was cleaned, stained with crystal violet for 10 minutes, and the remaining biomass was treated with anhydrous ethanol. The coatings were washed out and OD570 was detected by enzyme labeling.
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[[File:T--YAU-China--iGEM2019-BBa -1.png|500px|thumb|left|Figure 1. Effects of PelA and PslG on Biofilm]]
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[[File:T--YAU-China--iGEM2019-BBa -4.jpg|500px|thumb|none|Figure 1. Effects of PelA and PslG on Biofilm]]
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[[File:T--YAU-China--iGEM2019-BBa -3.png|500px|thumb|none|Figure 2. Crystal violet staining of biofilm treated with extracts from control group and three groups]]
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Latest revision as of 11:48, 15 October 2019


ptac+RBS+PelA+RBS+PslG

Biofilm is a complex bacterial community embedded in the extracellular matrix composed of proteins, extracellular DNA (eDNA) and extracellular polysaccharides. Pseudomonas aeruginosa PAO1 has two types of biofilm, Psl and Pel. This part is mainly used for encoding polysaccharide hydrolases PslG and PelA, and destroying the structure of biofilm.

Usage and Biology

YAU-China expressed the biofilm-damaged system in 2019, using extracts from each part of the system for experiments. After overnight induction of the control strain, two strains expressing single enzyme and two strains co-expressing two enzymes, four strains were collected by centrifugation and suspended in PBS buffer of equal volume. The supernatant was absorbed and added into the biofilm of Pseudomonas aeruginosa cultured on the pre-PVC plate. After incubation for a period of time, the PVC plate was cleaned, stained with crystal violet for 10 minutes, and the remaining biomass was treated with anhydrous ethanol. The coatings were washed out and OD570 was detected by enzyme labeling.

Figure 1. Effects of PelA and PslG on Biofilm


Figure 2. Crystal violet staining of biofilm treated with extracts from control group and three groups



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 769
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2922
    Illegal BglII site found at 3798
    Illegal BamHI site found at 1905
    Illegal BamHI site found at 2070
    Illegal XhoI site found at 3802
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 686
    Illegal NgoMIV site found at 710
    Illegal NgoMIV site found at 731
    Illegal NgoMIV site found at 832
    Illegal NgoMIV site found at 1058
    Illegal NgoMIV site found at 1766
    Illegal NgoMIV site found at 1815
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1652
    Illegal BsaI site found at 2278
    Illegal BsaI.rc site found at 1853