Difference between revisions of "Part:BBa K3739019"
 
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===Biology===
 
===Biology===
 
LamB is an anchor protein from Vibrio Species, which can anchor its passenger protein to the cell membrane. It has been widely used in cell-surface display. GFP is green fluorescent protein from jellyfish ''Aequorea Victoria'', which has been widely used as reporter for decades. GFP is fused at C terminal with LamB so that GFP may be displayed on the surface of VnDX.
 
LamB is an anchor protein from Vibrio Species, which can anchor its passenger protein to the cell membrane. It has been widely used in cell-surface display. GFP is green fluorescent protein from jellyfish ''Aequorea Victoria'', which has been widely used as reporter for decades. GFP is fused at C terminal with LamB so that GFP may be displayed on the surface of VnDX.
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===Usage===
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Here, we use BBa_3739019 to construct the expression system, which may achieve surface display on VnDX.
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===Characterization===
 
===Characterization===
 
====1. Identification====
 
====1. Identification====
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After receiving the synthesized DNA, PCR was done to certify that the plasmid was correct, and the experimental results were shown in figure1.
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The surface display system for ''Vibrio natriegens'' has never been reported before, so the performance of the surface display system of LamB needs to be verified preferentially. Surface display system bricks (BBa_K3739024, Fig. 1A) were assembled into plasmids, respectively, transformed into ''Vibrio natriegens'' through electroporation. After being verified by agarose gel electrophoresis, the plasmids have been transformed into ''Vibrio natriegens'' successfully (Fig. 1B).
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https://static.igem.org/mediawiki/parts/d/d3/T--XMU-China--K3739048.png
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::'''Fig. 1.''' Gene circuit and agarose gel electrophoresis. (A)  Gene circuit of ''LamB-GFP'' (BBa_K3739019). (B)Target bands of ''LamB-GFP'' (black arrow, 2300 bp).
  
  

Latest revision as of 19:43, 21 October 2021


LamB-GFP

This is an anchored proteins onto membranes through LamB and use GFP to comformation. We use BBa_K3739048 and GFP to verify LamB' s function which can anchor proteins onto membranes.

Biology

LamB is an anchor protein from Vibrio Species, which can anchor its passenger protein to the cell membrane. It has been widely used in cell-surface display. GFP is green fluorescent protein from jellyfish Aequorea Victoria, which has been widely used as reporter for decades. GFP is fused at C terminal with LamB so that GFP may be displayed on the surface of VnDX.

Usage

Here, we use BBa_3739019 to construct the expression system, which may achieve surface display on VnDX.

Characterization

1. Identification

The surface display system for Vibrio natriegens has never been reported before, so the performance of the surface display system of LamB needs to be verified preferentially. Surface display system bricks (BBa_K3739024, Fig. 1A) were assembled into plasmids, respectively, transformed into Vibrio natriegens through electroporation. After being verified by agarose gel electrophoresis, the plasmids have been transformed into Vibrio natriegens successfully (Fig. 1B). T--XMU-China--K3739048.png

Fig. 1. Gene circuit and agarose gel electrophoresis. (A) Gene circuit of LamB-GFP (BBa_K3739019). (B)Target bands of LamB-GFP (black arrow, 2300 bp).


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 528
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 676
    Illegal SapI.rc site found at 1246