Difference between revisions of "Part:BBa K3792001"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3792001 short</partinfo>
 
<partinfo>BBa_K3792001 short</partinfo>
  
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NodJ was expressed with NodC and NodI. We believe that NodJ is responsible for the transport of Chitin but we are not sure.  
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This part was lifted from the UniProt entry UniProtKB - P08720 [1]. This protein is from the Nodulation Operon (Nod) from Rhizobium leguminosarum bv. Viciae. This protein forms an ABC transporter cassette complex with Part BBa_K3792000 to transport Nodulation factors (Lipo-chito-oligomers or LCO’s) across the cell membrane to induce nodulation [2-5]. Nodulation is the process by which an organ is formed with a host plant to house the symbiotic bacterium.
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The 2021 FSU iGEM team has experimented with BBa_K2380000 and BBa_K3792001 to try and express chito-oligomers efficiently outside the cell membrane of E. Coli.
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The predicted structure of the NodIJ complex is available for download at https://2021.igem.org/Team:FSU/Model
  
 
To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan.
 
To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan.
  
 
Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture.
 
Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture.
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[1] https://www.uniprot.org/uniprot/P06755
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[2] Poinsot, Véréna, et al. “New Insights into Nod Factor Biosynthesis: Analyses of Chitooligomers and Lipo-Chitooligomers of Rhizobium Sp. IRBG74 Mutants.” Carbohydrate Research, vol. 434, 3 Nov. 2016, pp. 83–93., https://doi.org/10.1016/j.carres.2016.08.001.
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[3] Spaink, H P, et al. “Rhizobium NodI and NodJ Proteins Play a Role in the Efficiency of Secretion of Lipochitin Oligosaccharides.” Journal of Bacteriology, vol. 177, no. 21, Nov. 1995, pp. 6276–6281., https://doi.org/10.1128/jb.177.21.6276-6281.1995.
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[4] Mirdita, Milot, et al. “ColabFold - Making Protein Folding Accessible to All.” BioRxiv, Cold Spring Harbor Laboratory, 1 Jan. 2021, https://www.biorxiv.org/content/10.1101/2021.08.15.456425v1.
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[5] Hollenstein, Kaspar, et al. “Structure and Mechanism of ABC Transporter Proteins.” Current Opinion in Structural Biology, Elsevier Current Trends, 27 Aug. 2007, https://www.sciencedirect.com/science/article/pii/S0959440X07001029.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 03:00, 22 October 2021


NodJ

This part was lifted from the UniProt entry UniProtKB - P08720 [1]. This protein is from the Nodulation Operon (Nod) from Rhizobium leguminosarum bv. Viciae. This protein forms an ABC transporter cassette complex with Part BBa_K3792000 to transport Nodulation factors (Lipo-chito-oligomers or LCO’s) across the cell membrane to induce nodulation [2-5]. Nodulation is the process by which an organ is formed with a host plant to house the symbiotic bacterium.

The 2021 FSU iGEM team has experimented with BBa_K2380000 and BBa_K3792001 to try and express chito-oligomers efficiently outside the cell membrane of E. Coli.

The predicted structure of the NodIJ complex is available for download at https://2021.igem.org/Team:FSU/Model

To test if the cells were secreting chitin, The FSU 2021 team used the Chitosan Assay Kit (XAN-5126) available from Cell Biolabs. The manual for this kit contains the protocols for converting chitin to chitosan, along with a protocol for detecting chitosan.

Chitin from the cells was converted into chitosan by reacting with 12.5M NaOH at 95°C for 24 hours. The solution was then neutralized with acetic acid. The assay was then performed on the supernatant of the cell culture.

[1] https://www.uniprot.org/uniprot/P06755

[2] Poinsot, Véréna, et al. “New Insights into Nod Factor Biosynthesis: Analyses of Chitooligomers and Lipo-Chitooligomers of Rhizobium Sp. IRBG74 Mutants.” Carbohydrate Research, vol. 434, 3 Nov. 2016, pp. 83–93., https://doi.org/10.1016/j.carres.2016.08.001.

[3] Spaink, H P, et al. “Rhizobium NodI and NodJ Proteins Play a Role in the Efficiency of Secretion of Lipochitin Oligosaccharides.” Journal of Bacteriology, vol. 177, no. 21, Nov. 1995, pp. 6276–6281., https://doi.org/10.1128/jb.177.21.6276-6281.1995.

[4] Mirdita, Milot, et al. “ColabFold - Making Protein Folding Accessible to All.” BioRxiv, Cold Spring Harbor Laboratory, 1 Jan. 2021, https://www.biorxiv.org/content/10.1101/2021.08.15.456425v1.

[5] Hollenstein, Kaspar, et al. “Structure and Mechanism of ABC Transporter Proteins.” Current Opinion in Structural Biology, Elsevier Current Trends, 27 Aug. 2007, https://www.sciencedirect.com/science/article/pii/S0959440X07001029.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]