Difference between revisions of "Part:BBa K1139022"

 
 
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M13 is a filamentous phage that infects only F+ strains of E. coli, which does not kill the host cell. This biobrick is derived from M13mp18 phage vector, amplified by PCR. It includes 11 ORFs, M13 origin, a packaging sequence and Lac promoter. The promoter on the upstream of g2(gene 2) is deleted.The capsid is formed only when g2p(gene 2 protein) is expressed.
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We confirmed that M13 genome with two modifications related to our design kept plaque forming activity.  One is replacement of the promoter for G2p (gene 2 protein) with a constitutive promoter, PlacI<sup>q</sup> (<partinfo>BBa_I14032</partinfo>). The other is accommodation of pSB3K3 backbone. Even though the plasmid has two different types of replication origins, M13 origin and pSB3 origin, this plasmid (<partinfo>BBa_K1139020</partinfo>) formed plaque.  In contrast, construction intermediates without a promoter upstream of <i>g2p</i> coding sequence (Promoterless-M13 + Plac, Promoterless-M13 + Plac-GFP <partinfo>BBa_K1139022</partinfo>) could not form plaque.<br>
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[[Image:titech2013_parts_K1139022_exp_Fig1.jpg|thumb|left|400px|<b>Fig. 1.</b> PlacIQ-M13-Plac-GFP on pSB3 (<partinfo>BBa_K1139020</partinfo>)]]
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[[Image:titech2013_parts_K1139022_exp_Fig2.jpg|thumb|left|400px|<b>Fig. 2.</b> Promoterless-M13-Plac-GFP on pSB3 (<partinfo>BBa_K1139022</partinfo>)]]
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For more information, see [http://2013.igem.org/Team:Tokyo_Tech/Experiment/pSB-M13_Plasmid_Assay our work in Tokyo_Tech 2013 wiki].
  
 
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Latest revision as of 19:07, 27 October 2013

Promoterless-M13-Plac-GFP on pSB3

We confirmed that M13 genome with two modifications related to our design kept plaque forming activity. One is replacement of the promoter for G2p (gene 2 protein) with a constitutive promoter, PlacIq (BBa_I14032). The other is accommodation of pSB3K3 backbone. Even though the plasmid has two different types of replication origins, M13 origin and pSB3 origin, this plasmid (BBa_K1139020) formed plaque. In contrast, construction intermediates without a promoter upstream of g2p coding sequence (Promoterless-M13 + Plac, Promoterless-M13 + Plac-GFP BBa_K1139022) could not form plaque.

Fig. 1. PlacIQ-M13-Plac-GFP on pSB3 (BBa_K1139020)
Fig. 2. Promoterless-M13-Plac-GFP on pSB3 (BBa_K1139022)


















For more information, see [http://2013.igem.org/Team:Tokyo_Tech/Experiment/pSB-M13_Plasmid_Assay our work in Tokyo_Tech 2013 wiki].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 100
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 6026
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 7287