Difference between revisions of "Part:BBa K2387060"

Revision as of 14:18, 19 October 2017

Double tagged ISG64 of T.b. gambiense

This part contains the extracellular domain (domain 24-365) of the invariant surface glycoprotein 64 (ISG64) of Trypanosoma brucei gambiense: BBa_K2387058. This parasite causes African Sleeping Sickness (Human African Trypanosomiasis). ISG64 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
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 811

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2387060 short</partinfo>
-This part contains the extracellular domain (domain 24-365) of the invariant surface glycoprotein 64 (ISG64) of Trypanosoma brucei gambiense: [https://parts.igem.org/Part:BBa_K2387058 BBa_K2387058]. This parasite causes African Sleeping Sickness (Human African Trypanosomiasis). ISG64 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 24-365) of the invariant surface glycoprotein 64 (ISG64) of Trypanosoma brucei gambiense: [https://parts.igem.org/Part:BBa_K2387058 BBa_K2387058]. This parasite causes African Sleeping Sickness (Human African Trypanosomiasis). ISG64 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.