Part:BBa_K4195007
ttpA-his
Biology
TTPA is the phage tail tubular protein A of podophage 7. TTPA can interact with Vp0980, which acts as the receptor of TTPA on the surface of Vibrio parahaemolyticus. TTPA’s binding to Vp0980 mediates phage absorption and subsequent bacterial lysis (1).
Usage and design
A His-Tag (6×His) was added to the C-terminal of TTPA for purifying this protein. Arabinose-inducible system was used in the expression circuit at pSB1C3 then composite part BBa_K4195111 was obtained. We transformed the plasmid into E. coli SHuffle T7 to obtain the protein for further biochemical characterizations.
Characterization
Identification
When constructing this circuit, colony PCR and gene sequencing were used to verify that the transformatants were correct. Target bands (764 bp) can be observed at the position around 750 bp (Fig. 1).
Fig. 1 DNA gel electrophoresis of the colony PCR products of BBa_K4195007_pSB1C3.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 520
- 1000COMPATIBLE WITH RFC[1000]
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