Difference between revisions of "Part:BBa K4195007"

 
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<partinfo>BBa_K4195007 short</partinfo>
 
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===Biology===
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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'').
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===Usage and design===
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===Usage and Biology===
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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.
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===Characterization===
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====Identification====
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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).
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4195007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4195007 SequenceAndFeatures</partinfo>
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Revision as of 12:50, 10 October 2022


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). 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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 520
  • 1000
    COMPATIBLE WITH RFC[1000]