Composite

Part:BBa_K4268018

Designed by: J. Aubrey, J. Alvarenga, L. Buchanan, J. Reyes, D. Yashinski     Group: iGEM22_SUNY_Oneonta   (2022-09-30)


Biotin Tagged Capsid and Core Multi-Transcriptional Unit

Usage and Biology

S-TIP 37 is a T7-like cyanophage that infects its host via a lytic life cycle (Shitrit et al., 2021). T7-like phages are characterized by a complex symmetrical capsid structure, which includes an icosahedral head that houses the phage's genome, a neck region that facilitates DNA delivery into a host, and six tail fibers used for attachment to its host. (Raytcheva et al., 2011).

Figure 1: A labeled visual detailing the various structures of a T7-like phage (Kemp et al., 2005)


This part belongs to a collection that codes for a "ghost" phage. This ghost phage is a capsid-only, empty viral shell that could be modified to immobilize Cyanobacteria recognized by the viral tail fibers or used with modification to deliver substances to a chassis Cyanobacteria.

However, it will infect Synechococcus sp WH 8109, the cyanobacteria strain that is the natural host of S-TIP 37. Further modeling will be needed to determine if the "ghost" phage could effectively target other strains of Cyanobacteria that are used in synthetic biology, such as Synechococcus sp PCC 11901.

The Biotin Tagged Capsid and Core Multi-Transcriptional Unit (MTU) codes for the putative proteins that make the icosahedral head (Internal Core, Biotin Tagged Capsid, and Capsid Assembly Protein) of S-TIP 37 and a dummy part to facilitate assembly into an MTU. This MTU is intended to be used with the Neck and Tail MTU (part BBa_K4268016) to create an S-TIP 37 ghost phage capsid. This phage will contain a biotin-tagged icosahedral head region, which can be used to detect or immobilize the ghost phage for downstream applications in conjunction with streptavidin.

The Capsid Protein in other T7 viruses serves to form the icosahedral head of the virus which normally houses the viral genome, while the Capsid Assembly Protein and core region ensure proper assembly of the head. In addition, the internal core stabilizes the packaged DNA inside the head.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 3806
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 3806
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 5170
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 3806
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 3806
    Illegal NgoMIV site found at 3446
    Illegal NgoMIV site found at 3585
    Illegal NgoMIV site found at 4541
  • 1000
    COMPATIBLE WITH RFC[1000]


References

Shitrit D, Hackl T, Laurenceau R, Raho N, Carlson MCG, Sabehi G, Schwartz DA, Chisholm SW, Lindell D. Genetic engineering of marine cyanophages reveals integration but not lysogeny in T7-like cyanophages. ISME J. 2022 Feb;16(2):488-499. doi: 10.1038/s41396-021-01085-8. Epub 2021 Aug 24. PMID: 34429521; PMCID: PMC8776855.

Raytcheva DA, Haase-Pettingell C, Piret JM, King JA. Intracellular assembly of cyanophage Syn5 proceeds through a scaffold-containing procapsid. J Virol. 2011 Mar;85(5):2406-15. doi: 10.1128/JVI.01601-10. Epub 2010 Dec 22. PMID: 21177804; PMCID: PMC3067778.

Kemp P, Garcia LR, Molineux IJ. Changes in bacteriophage T7 virion structure at the initiation of infection. Virology. 2005 Sep 30;340(2):307-17. doi: 10.1016/j.virol.2005.06.039. PMID: 16054667

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