Difference between revisions of "Part:BBa K3286106"
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<p> To prove chitin binding, a chitin binding assay was conducted. In a 2 ml microfuge tube, 500 mg of chitin solution (10 mg/ml) was added, together with 50 ug of protein in solution. The final volume was adjusted, using MQ, to 500 ul. Then, the tubes were incubated at room temperature in a shaking block at 600 rpm. After 1 hour, the tubes were centrifuged for 3 minutes at 13000 x g. Then, the supernatant was used to perform a Bradford assay with.
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In this experiment, 4 different samples were used (triplicates): PD1764sh with chitin, PD1764sh, BSA and BSA with chitin.
 
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Revision as of 14:38, 21 October 2019


Shorter version of PD1764

This protein is a shorter (sh) version of BBa_K3286105. Due to isolation troubles, the first 20 amino acids have been removed from this protein. The part contains a chitin-binding domain that is essential for chitin binding capacities [1].

[1] F. Labroussaa, A. R. Zeilinger, and R. P. P. Almeida, “Blocking the Transmission of a Noncirculative Vector-Borne Plant Pathogenic Bacterium,” Mol. Plant-Microbe Interact., vol. 29, no. 7, pp. 535–544, Jul. 2016.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 787
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 294
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Functional Characterization

Figure 1: Protein concentration in supernatant in ug/ml. 55% of PD1764sh has bound to chitin in this assay. The influence of chitin on protein concentration in general is negligible, as the BSA sample indicates

To prove chitin binding, a chitin binding assay was conducted. In a 2 ml microfuge tube, 500 mg of chitin solution (10 mg/ml) was added, together with 50 ug of protein in solution. The final volume was adjusted, using MQ, to 500 ul. Then, the tubes were incubated at room temperature in a shaking block at 600 rpm. After 1 hour, the tubes were centrifuged for 3 minutes at 13000 x g. Then, the supernatant was used to perform a Bradford assay with. In this experiment, 4 different samples were used (triplicates): PD1764sh with chitin, PD1764sh, BSA and BSA with chitin.