Part:BBa_K5048068:Experience

Experiments

This part acts as an adhesin construct that can display protein LAP on the outer membrane of E. coli and adhere to HSP60.

The overall experiments we conducted on BBa_K5048058 include:

1. Verification of PCR products
2. Verification of transformation of the plasmid

Resultss

Verification of PCR products

For the Neae-LAP plasmid, although the PCR primers lack specificity, we still obtain the targeted band(Fig 1) and implement purification to acquire the targeted segments.

Fig 1 The input for T4 ligase.
The target gene segments are marked in the red box. Segment LAP is marked in the red box below while the segment Neae backbone is marked in the upper box. Neae backbone: 7259bp; LAP: 2649bp

Verification of the transformation

For pET-28a-[Neae-LAP], we ultimately successfully transferred the plasmid into E. coli BL21(DE3) and had a corresponding sequencing result(Fig 2).

Fig 2 Neae-LAP successfully transferred into E. coli BL21(DE3)
A. The colony PCR for E. coli BL21(DE3) strains. Neae-LAP 3 has the segment corresponding to the estimated DNA length. Neae-LAP: 4831bp; B. The sequencing result for Neae-LAP in E. coli BL21(DE3), demonstrates the successful transformation.

Applications of BBa_K5048068

A part of Adhesion Construct that can display protein LAP on the outer membrane of E. coli and adhere to HSP60 in engineered bacteria and small intestine.

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