Coding

Part:BBa_K112805

Designed by: Jin Huh   Group: iGEM08_UC_Berkeley   (2008-10-30)
Revision as of 01:53, 10 October 2023 by LHL scau (Talk | contribs) (Characterized by SCAU-China 2023)


[T4 holin]

Holins from T4 bacteriophage assemble together to form pores on inner membrane of bacteria allowing lysozyme to reach periplasm and degrade peptidoglycan layer.

Group: (Michigan 2017) Author: (Aaron Renberg) Summary: We improved this part by optimizing the codons for translation in E. coli using IDT’s codon optimization tool, and by eliminating the illegal XbaI site that Imperial College London’s 2011 team found, making it much easier for future iGEM teams to use. The changes we made were T358C, T556C, T563C, and T571C. Additionally, we constructed three different versions (of varying promoter strength) of a temperature controlled kill switch using holin, endolysin and antiholin. Link: https://parts.igem.org/Part:BBa_K2301000

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]


Characterized by CAFA_China 2022

  • We constucted a gene circuit include lacZ gene (BBa_I732019) and T4 lysis Device.
T4 lysis Device and beta-galactosidase synthesis

  • The experimental result shows that the OD600 of recombinant cells with pBAD-lacZ-T4 lysis gene circuit reduced significantly by 2-3 times than non-recombinant cells after induced by different concentrations of arabinose.
Figure: The result of T4 lysis Device after induced by different concentrations of arabinose. (pSB1C3: non-recombinant DH10B without arabinose induction; pSB1C3-pBAD-lacZ-T4 lysis: recombinant DH10B without arabinose induction and with different concentrations of arabinose).


Characterized by SCAU-China 2023

What have we done?

pLas was from BBa_K2967001[1] We introduced it to achieve control over drug concentration through bacterial density.

To achieve control over toxicant concentration, we introduced components labeled with BBa_K4632016 [2] and BBa_K112806[3] to create the T4-T4 lysis device. BBa_K4632016 was originally from BBa_K112805[4]. Its Codon was optimized for Escherichia coli expression.

We characterized the component to demonstrate its effectiveness, as detailed in the construction and characterization section.

[edit]
Categories
Parameters
None