Difference between revisions of "Part:BBa K3739051"

Revision as of 18:05, 21 October 2021

J23100-B0030-LamB-LC1KR-2-GFP-B0010

We anchor LCI-KR2 protein onto membranes through LamB to stick the engineered bacteria on the polypropylene. We use BBa_K3739051 to construct the expression system and anchor LCI-KR2 on the surface of VnDX.

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. LCI-KR2, A mutant of a peptide called LCI from Bacillus subtilis, can sticks to polypropylene strongly. LCI-KR2 is fused at C terminal with LamB so that LCI-KR2 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 J23100-B0030-LamB-LC1KR-2-GFP-B0010 (BBa_K3739051). (B)Target bands of J23100-B0030-LamB-LC1KR-2-GFP-B0010 (black arrow, 2300 bp).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 592
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 740
Illegal SapI.rc site found at 1496

@@ Line 12: / Line 12: @@
 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).
+::'''Fig. 1.''' Gene circuit and agarose gel electrophoresis. (A)  Gene circuit of J23100-B0030-LamB-LC1KR-2-GFP-B0010 (BBa_K3739051). (B)Target bands of J23100-B0030-LamB-LC1KR-2-GFP-B0010 (black arrow, 2300 bp).
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