Coding

Part:BBa_K3552029

Designed by: Liu Xibei   Group: iGEM20_LINKS_China   (2020-10-25)
Revision as of 11:54, 25 October 2020 by Registry (Talk | contribs)

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gspO

GspO plays a role in type II pseudopili formation by removing the leader sequence from substrate proteins and subsequently methylating the alpha-amino group of the newly exposed N-terminal phenylalanine. Substrates include proteins required for biogenesis of the type II general secretory apparatus. This part is in the part collection where we have 12 genes that code for the base generator of pilA.

The part collection includes: Parts that are different kinds of type 4 pilus: BBa_K3552000 BBa_K3552001 BBa_K3552002. Parts that are the generator of the type 4 pilus: BBa_K3552003 BBa_K3552004 BBa_K3552005 BBa_K3552006 BBa_K3552007 BBa_K3552008 BBa_K3552018 BBa_K3552019 BBa_K3552020 BBa_K3552021 BBa_K3552022 BBa_K3552023 BBa_K3552024 BBa_K3552025 BBa_K3552026 BBa_K3552027 BBa_K3552028 BBa_K3552029. Parts that are a complete circuit: BBa_K3552009 BBa_K3552010 BBa_K3552011 BBa_K3552012.

Our part collection can instruct other teams to designed new rechargeable pilus and substitution of different major pilin.

Usage and Biology: The gene gspO, one of the two prepilin peptidase genes in E.coli K-12, is the last gene of the gsp operon encoding the chitinase-specific type 2 secretion system. In EHEC, almost all gsp genes have been lost during evolution, leaving only the gspO fragment encoding the prepilin peptidase catalytic domain. Additionally, EHEC strains have the gspO homologue etpO within a plsmid-encoded type 2 secretion system cluster. This gene is also a part of the generator of the major pilin PilA.

Characterization: We tried to uncoupled all the genes into single ones and we already separated them. We mutated the sequence and codon optimization of the repeated sequence from two genes to prevent recombination. In the future we are going to link all single genes to the same promotor and reconstructed all genes into a complete circuit to see any difference comparing to the initial combined genes.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 18


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