Coding

Part:BBa_K4905005

Designed by: Merel van den Bosch   Group: iGEM23_TU-Eindhoven   (2023-07-23)
Revision as of 13:24, 4 September 2023 by VeerleJegers (Talk | contribs)

Leucine zipper Z2

Information

Basic-region Leucine zippers (bZIPs) are alpha-helical domains with a repeating unit “abcdefg”. In this unit, the positions “a” and “d” consist of a hydrophobic residue and the ”e” and “g” positions consist of charged residues. The zippers form an alpha-helix and their charged residues form ion pairs between helices, causing them to associate [1].

Individual bZIP proteins can form homodimers or heterodimers with other bZIP proteins with a slightly different sequence[1]. Not all bZIP proteins can dimerize with each other, some will only homodimerize with the same bZIP protein. Others will only heterodimerize with specific other bZIP proteins [2].

In cells, the main function of bZIPs is that they work as transcription factor, where the homodimer or heterodimer will form at the promotor regions of target genes [3]. This makes leucine zippers a large family of transcription factors, where each member has a preference for a specific DNA sequence [2].

The TU-Eindhoven 2023 team used this part in a composite part for the formation of a hydrogel in E. coli. Two different bZIP proteins are made, which will form a heterodimer. This part is one of these bZIP proteins.

Characterization

We ran an electrophoresis gel with the plasmid, digested with enzymes AcuI and BglI. The band of Z2 is shown in slot 5 in the gel. A 1 kb ladder (L) was used. The remaining slots were used for different parts. The bands formed as expected.

  1. A120: BseRI + BglI, with CIP
  2. Z1.1: Acul + BglI, without CIP
  3. A100: BseRI + BglI, with CIP
  4. Z1.2: Acul + BglI, without CIP
  5. Z2: Acul + BglI, without CIP

caption

References

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

[2] Hakoshima, T. (n.d.). Leucine Zippers. https://doi.org/10.1038/npg.els.0005049

[3] Seldeen, K. L., McDonald, C. B., Deegan, B. J., Bhat, V., & Farooq, A. (2010). Dissecting the Role of Leucine Zippers in the Binding of bZIP Domains of Jun Transcription Factor to DNA. Biochemical and Biophysical Research Communications, 394(4), 1030. https://doi.org/10.1016/J.BBRC.2010.03.116 caption

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