Coding

Part:BBa_K3739024

Designed by: Han Ziyan   Group: iGEM21_XMU-China   (2021-09-08)
Revision as of 19:17, 21 October 2021 by Teslwang (Talk | contribs)


LamB-LC1KR2-GFP

We anchor LCIKR-2 protein onto membranes through LamB to stick the engineered bacteria on the polypropylene. We use BBa_K3739051 to construct the expression system and anchor LCIKR-2 on the surface of VnDX.

Usage and 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. LCIKR-2, A mutant of a peptide called LCI from Bacillus subtilis, can sticks to polypropylene strongly. LCIKR-2 is fused at C terminal with LamB so that LCIKR-2 can be displayed on the surface of VnDX.

Characterization

1. Identification

After receiving the synthesized DNA, PCR was done to certify that the plasmid was correct, and the experimental results were shown in figure1.

Fig. 1. Gene circuit and agarose gel electrophoresis. (A) Gene circuit of LamB-LCIKR-2-GFP (BBa_K3739024). (B)Target bands of LamB-LCIKR-2-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 1432


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