Difference between revisions of "Part:BBa K4375018"

 
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<partinfo>BBa_K4375018 short</partinfo>
 
<partinfo>BBa_K4375018 short</partinfo>
  
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The MiniNano is a compact construct meant for extracellular protein (like tumour-specific receptor, antigen, etc.) detection via fluorescence. This protein has 3 domains. (1) INPNC surface display module (INPNC codes for N- and C- terminal domain of Ice Nucleation Protein (INP) from Pseudomonas syringae, and it is used for displaying proteins on bacteria's outer membrane (BBa_K4375007)). It is linked to (by a flexible GS-linker, BBa_K4375010)
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This construction consists of 4 described parts. These are:  Codon optimalised INPNC for Surface Display (BBa_K4375007), flexible GS-linker (BBa_K4375010),miRFP670 nano (BBa_K4375008), J23101* (BBa_K4375003) . They allow the bacterium to be expelled to the tumour surface, thus allowing its visualisation and colonisation on the tumour surface.
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) (2) a small, near-infrared emitting fluorescent protein, miRFP670 nano (BBa_K4375008)
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), which is connected to an anti-EGFR specific Nanobody, 7D12, via llama-derived igA hinge region linker (BBa_K4375011). Moreover, it contains a Myc epitope tag, so the specific interaction can be detected in another way. This complex detection device is expressed under the constitutive promoter J23101* (BBa_K4375003)  
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The Nanobody in the construct can be easily swapped with other protein-domains, as its sequence is framed by BamHI and SphI restriction sites.
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===Usage and Biology===
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==Usage and Biology==
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The MiniNano is a compact construct meant for extracellular protein (like tumour-specific receptor, antigen, etc.) detection via fluorescence. This protein has 3 domains. (1) INPNC surface display module (INPNC codes for N- and C- terminal domain of Ice Nucleation Protein (INP) from Pseudomonas syringae, and it is used for displaying proteins on bacteria's outer membrane (BBa_K4375007)). It is linked to (by a flexible GS-linker, BBa_K4375010) ) (2) a small, near-infrared emitting fluorescent protein, miRFP670 nano (BBa_K4375008) ), which is connected to an anti-EGFR specific Nanobody, 7D12, via llama-derived IgA hinge region linker (BBa_K4375011). Moreover, it contains a Myc epitope tag, so the specific interaction can be detected in another way. This complex detection device is expressed under the constitutive promoter J23101* (BBa_K4375003) The Nanobody in the construct can be easily swapped with other protein-domains, as its sequence is framed by BamHI and SphI restriction sites.
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Revision as of 09:39, 7 October 2022


MiniNano for bacterial surface display

This construction consists of 4 described parts. These are: Codon optimalised INPNC for Surface Display (BBa_K4375007), flexible GS-linker (BBa_K4375010),miRFP670 nano (BBa_K4375008), J23101* (BBa_K4375003) . They allow the bacterium to be expelled to the tumour surface, thus allowing its visualisation and colonisation on the tumour surface.


Usage and Biology

The MiniNano is a compact construct meant for extracellular protein (like tumour-specific receptor, antigen, etc.) detection via fluorescence. This protein has 3 domains. (1) INPNC surface display module (INPNC codes for N- and C- terminal domain of Ice Nucleation Protein (INP) from Pseudomonas syringae, and it is used for displaying proteins on bacteria's outer membrane (BBa_K4375007)). It is linked to (by a flexible GS-linker, BBa_K4375010) ) (2) a small, near-infrared emitting fluorescent protein, miRFP670 nano (BBa_K4375008) ), which is connected to an anti-EGFR specific Nanobody, 7D12, via llama-derived IgA hinge region linker (BBa_K4375011). Moreover, it contains a Myc epitope tag, so the specific interaction can be detected in another way. This complex detection device is expressed under the constitutive promoter J23101* (BBa_K4375003) The Nanobody in the construct can be easily swapped with other protein-domains, as its sequence is framed by BamHI and SphI restriction sites.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1055
    Illegal PstI site found at 1313
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 29
    Illegal PstI site found at 1055
    Illegal PstI site found at 1313
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1055
    Illegal PstI site found at 1313
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1055
    Illegal PstI site found at 1313
    Illegal NgoMIV site found at 118
    Illegal NgoMIV site found at 451
    Illegal AgeI site found at 869
    Illegal AgeI site found at 1523
    Illegal AgeI site found at 1655
  • 1000
    COMPATIBLE WITH RFC[1000]