Difference between revisions of "Part:BBa K2387061"
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This part contains the extracellular domain (domain 21-385) of the invariant surface glycoprotein 65 (ISG65) of Trypanosoma brucei gambiense:  [https://parts.igem.org/Part:BBa_K2387059 BBa_K2387059]. This parasite causes African Sleeping Sickness (Human African Trypanosomiasis). ISG65 is a surface protein and thus antigen of this organism. It is double tagged, having an N-terminal Strep-tag II and C-terminal 10xHis tag for the convenience of purification. Moreover, because of two distinct cleavage sites, the tags can be cleaved of if necessary (see [https://parts.igem.org/Part:BBa_K2387056 BBa_K2387056] and [https://parts.igem.org/Part:BBa_K2387057 BBa_K2387057]).
 
This part contains the extracellular domain (domain 21-385) of the invariant surface glycoprotein 65 (ISG65) of Trypanosoma brucei gambiense:  [https://parts.igem.org/Part:BBa_K2387059 BBa_K2387059]. This parasite causes African Sleeping Sickness (Human African Trypanosomiasis). ISG65 is a surface protein and thus antigen of this organism. It is double tagged, having an N-terminal Strep-tag II and C-terminal 10xHis tag for the convenience of purification. Moreover, because of two distinct cleavage sites, the tags can be cleaved of if necessary (see [https://parts.igem.org/Part:BBa_K2387056 BBa_K2387056] and [https://parts.igem.org/Part:BBa_K2387057 BBa_K2387057]).
 +
 
By only taking the extracellular domain, protein expression and purification are simplified since a soluble protein is formed. Note that this DNA originates from parasitic genomic DNA, and codon optimization has not been performed. Hence, it is advised to use an E. coli strain optimized for this.
 
By only taking the extracellular domain, protein expression and purification are simplified since a soluble protein is formed. Note that this DNA originates from parasitic genomic DNA, and codon optimization has not been performed. Hence, it is advised to use an E. coli strain optimized for this.
 +
 
This protein can be used for, for example, a proof-of-principle for the binding to or detection of African Sleeping Sickness.
 
This protein can be used for, for example, a proof-of-principle for the binding to or detection of African Sleeping Sickness.
  

Revision as of 14:19, 19 October 2017


Double tagged ISG65 of T.b. gambiense

This part contains the extracellular domain (domain 21-385) of the invariant surface glycoprotein 65 (ISG65) of Trypanosoma brucei gambiense: BBa_K2387059. This parasite causes African Sleeping Sickness (Human African Trypanosomiasis). ISG65 is a surface protein and thus antigen of this organism. It is double tagged, having an N-terminal Strep-tag II and C-terminal 10xHis tag for the convenience of purification. Moreover, because of two distinct cleavage sites, the tags can be cleaved of if necessary (see BBa_K2387056 and BBa_K2387057).

By only taking the extracellular domain, protein expression and purification are simplified since a soluble protein is formed. Note that this DNA originates from parasitic genomic DNA, and codon optimization has not been performed. Hence, it is advised to use an E. coli strain optimized for this.

This protein can be used for, for example, a proof-of-principle for the binding to or detection of African Sleeping Sickness.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 823
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]