Composite

Part:BBa_K4905008

Designed by: Merel van den Bosch   Group: iGEM23_TU-Eindhoven   (2023-08-14)


Elastin-like polypeptide triblock with leucine zippers

Information

This part is made up of the basic parts: two times Leucine zipper Z2 (BBa_K4905005), and two times Elastin-Like Polypeptide (ELP) sequence A[60]I[60] (BBa_K4905001). This results in the sequence Z2-I[60]-A[120]-I[60]-Z2. With A[5] the sequence (VPGAG[3]VPGGG[2]), since there are five VPGXG repeats, and I the sequence (VPGIG). The numbers indicate the number of repeats of these sequences. This construct was used by the TU Eindhoven 2023 team to form a hydrogel outside as well as inside E.coli BL21 cells. A schematic overview of this is shown in figure 1.

Figure 1 | Schematic overview of the sequence of this construct. (VPGAG[3]VPGGG[2]) is from now on referred to as A[5] and VPGIG is referred to as I.

General applications

This specific part was used as a control protein for part BBa_K4905006 . This part has two Leucine zippers that are not complementary, while part BBa_K4905006 has complementary Leucine zippers. By also testing the part with non-complementary zippers, it was possible to find out their function. More lab results can be read on the part page of BBa_K4905006 .

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1982
    Illegal EcoRI site found at 3908
    Illegal XbaI site found at 99
    Illegal XbaI site found at 2025
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1982
    Illegal EcoRI site found at 3908
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1982
    Illegal EcoRI site found at 3908
    Illegal XhoI site found at 1999
    Illegal XhoI site found at 3925
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1982
    Illegal EcoRI site found at 3908
    Illegal XbaI site found at 99
    Illegal XbaI site found at 2025
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1982
    Illegal EcoRI site found at 3908
    Illegal XbaI site found at 99
    Illegal XbaI site found at 2025
    Illegal NgoMIV site found at 156
    Illegal NgoMIV site found at 336
    Illegal NgoMIV site found at 426
    Illegal NgoMIV site found at 606
    Illegal NgoMIV site found at 2082
    Illegal NgoMIV site found at 2262
    Illegal NgoMIV site found at 2352
  • 1000
    COMPATIBLE WITH RFC[1000]

References

[1] Alber, T. (1992). Structure of the leucine zipper. Current Opinion in Genetics and Development, 2, 205–210

[2] Christensen, T., Hassouneh, W., Trabbic-Carlson, K., & Chilkoti, A. (2023). Predicting Transition Temperatures of Elastin-Like Polypeptide Fusion Proteins. https://doi.org/10.1021/bm400167h

[3] Despanie, J., Dhandhukia, J. P., Hamm-Alvarez, S. F., & MacKay, J. A. (2016). Elastin-like polypeptides: Therapeutic applications for an emerging class of nanomedicines. Journal of Controlled Release, 240, 93–108. https://doi.org/10.1016/j.jconrel.2015.11.010

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